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HoC_dynamic_5_shot_test

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HoC_dynamic_5_shot0

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cellular senescence is considered as a tumor suppressive mechanism . Recent evidence indicates however that senescent cells secrete various growth factors and cytokines , some of which may paradoxically promote cancer progression . This phenomenon termed senescence-associated secretory phenotype ( SASP ) must be inhibited in order for anti-proliferative agents to be effective . The present study was designed to determine whether the β-catenin destruction complex ( BCDC ) , known to integrate the action of various growth factors and cytokines , would represent a suitable target to inhibit the activity of SASP components . For this , we carried out experiments to determine the effect of drug-induced senescence on secretion of SASP , β-catenin transactivation , and the relationship between these processes . Moreover , genetic and pharmacological approaches were used to define the implication of BCDC in mediating the effects of SASP components on cell migration and resistance to drugs . The findings indicate that drug-induced senescence was associated with expression of various Wnt ligands in addition to previously known SASP components . Beta catenin transactivation and expression of genes implicated in epithelial-mesenchymal transition ( EMT ) also increased in response to drug-induced SASP . These effects were prevented by Pyrvinium , a recently described activator of BCDC . Pyrvinium also suppressed the effects of SASP on cell migration and resistance to doxorubicin . Together , these findings provide insights on the potential role of BCDC in mediating the effects of drug-induced SASP on cancer cell invasion and resistance to therapy , and suggest that targeting this pathway may represent an effective approach to enhance the activity of current and prospective anti-cancer therapeutics . OUTPUT: enabling replicative immortality;activating invasion and metastasis INPUT: Ovarian cancer-related angiogenesis is a complex process orchestrated by many positive and negative regulators . Many growth factors are involved in the development of the tumor-associated vasculature , and from these , endocrine gland-derived vascular endothelial growth factor ( EG-VEGF ) seems to play a crucial role . EG-VEGF is the first organ-specific angiogenic factor and its effects are restricted to the endothelial cells of the endocrine glands . Although EG-VEGF was detected in both normal and neoplastic ovaries , its clinical significance remains controversial . In the present study , we analyzed 30 patients with epithelial ovarian cancer , and the immunohistochemical expression of EG-VEGF was compared with the conventional clinico-pathological parameters of prognosis . Neoplastic cells of the ovarian carcinoma expressed EG-VEGF in 73.33% of the cases , as a cytoplasmic granular product of reaction . We found a strong correlation between the expression of EG-VEGF at protein level and tumor stage , grade , and microscopic type . The expression of EG-VEGF was found in patients with stage III and IV , but not in stage II . The majority of serous adenocarcinoma , half of the cases with clear cell carcinoma and two cases with endometrioid carcinoma showed definite expression in tumor cells . No positive reaction was found in the cases with mucinous carcinoma . Our results showed that EG-VEGF expression is an indicator not only of the advanced stage , but also of ovarian cancer progression . Based on these data , we concluded that EG-VEGF expression in tumor cells of the epithelial ovarian cancer is a good marker of unfavorable prognosis and could be an attractive therapeutic target in patients with advanced-stage tumors , refractory conventional chemotherapy . OUTPUT: inducing angiogenesis INPUT: Epidermal growth factor receptor-tyrosine kinase inhibitors ( EGFR-TKIs ) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor ( EGFR ) gene . On the other hand , some lung cancer patients with wild type EGFR also respond to EGFR-TKIs , suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells . However , the effect of EGFR-TKIs on host microenvironments is largely unknown . A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells . This model was used to investigate the therapeutic efficacy of erlotinib , an EGFR-TKI , on multiple organ metastases induced by human small cell lung cancer cells ( SBC-5 cells ) that did not express EGFR . Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro , it significantly suppressed bone and lung metastases in vivo , but not liver metastases . An immunohistochemical analysis revealed that , erlotinib significantly suppressed the number of osteoclasts in bone metastases , whereas no difference was seen in microvessel density . Moreover , erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line ( MC3T3-E1 cells ) . These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells . OUTPUT: activating invasion and metastasis;inducing angiogenesis;sustaining proliferative signaling INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Breast cancer incidence is increased in women receiving menopausal hormone therapy with estrogen plus progestin but not with estrogen alone . The use of a tissue-selective estrogen complex ( TSEC ) has been proposed as a novel menopausal hormone therapy strategy to eliminate the requirement for a progestogen . Combination of bazedoxifene ( BZA ) with conjugated estrogens ( CEs ) , the first TSEC , has shown beneficial effects . Whether it would exert antiestrogenic effects on breast cancer is not clear . To address this issue , we compared estradiol ( E(2) ) and CE alone on proliferation and apoptosis in MCF-7 breast cancer cells . CE stimulated growth of MCF-7 cells at a peak concentration 10-fold higher than required for E(2) . Both CE and E(2) alone increased DNA synthesis and reduced apoptosis with activation of MAPK , Akt , and p70S6K and up-regulation of antiapoptotic factors survivin , Bcl-2 , and X-linked inhibitor of apoptosis protein , These effects could be completely blocked by BZA . Gene expression studies demonstrated that CE and E(2) were equally potent on expression of cMyc , pS2 , and WNT1 inducible signaling pathway protein 2 , whereas the stimulatory effects of CE on progesterone receptor and amphiregulin expression were weaker than E(2) . BZA effectively blocked each of these effects and showed no estrogen agonistic effects when used alone . Our results indicate that the stimulatory effects of E(2) or CE on breast cancer cells could be completely abrogated by BZA . These studies imply that the CE/BZA , TSEC , exerts antiestrogenic effects on breast cancer cells and might block the growth of occult breast neoplasms in postmenopausal women , resulting in an overall reduction in tumor incidence . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: PURPOSE We recently reported that overexpression of epidermal growth factor receptor ( EGFR ) positively correlated with radioresistance of murine carcinomas . Because cyclin D1 is a downstream sensor of EGFR activation , the present study investigated whether a relationship exists between the extent of cyclin D1 expression and in vivo radiocurability of murine tumors . We further investigated the influence of radiation on cyclin D1 expression and the expression of p27 , an inhibitor of the cyclin D1 downstream pathway , as well as the relationship of these molecular determinants to cell proliferation and induced apoptosis in tumors exposed to radiation . METHODS AND MATERIALS Cyclin D1 expression was assayed in nine carcinomas syngeneic to C3Hf/Kam mice using Western blot analysis . These tumors greatly differed in their radioresponse as assessed by TCD(50) . The expression of cyclin D1 and p27 proteins was determined by Western blotting . Cell proliferative activity in tumors was determined by proliferating cell nuclear antigen ( PCNA ) immunochemistry . The effect of irradiation on the expression of cyclin D1 or p27 proteins and on PCNA positivity was determined in the radiosensitive OCa-I and in the radioresistant SCC-VII tumors . RESULTS Cyclin D1 expression varied among tumors by 40-fold , and its magnitude positively correlated with poorer tumor radioresponse ( higher TCD(50) values ) . The level of cyclin D1 expression paralleled that of EGFR . A 15-Gy dose reduced constitutive expression of cyclin D1 in the radiosensitive OCa-I tumors , but had no influence on expression of cyclin D1 in the radioresistant SCC-VII tumors . In contrast , 15 Gy increased the expression of p27 in radiosensitive tumors and reduced it in radioresistant tumors . Radiation induced no significant apoptosis or change in the percentage of PCNA-positive ( proliferating ) cells in SCC-VII tumors with high cyclin D1 levels , but it induced significant apoptosis and a decrease in the percentage of proliferating cells in OCa-I tumors with low cyclin D1 expression . CONCLUSION Our findings show a positive correlation between cyclin D1 expression and tumor radioresistance . The expression of cyclin D1 and p27 was modified by radiation and was associated with cellular response to radiation , but this depended on the pretreatment level of cyclin D1 expression . These findings may have important clinical implications : The pretreatment assessment of cyclin D1 expression could serve as a useful predictor of radiotherapy outcome and assist in selecting an effective treatment modality . OUTPUT:

sustaining proliferative signaling;evading growth suppressors;resisting cell death

HoC_dynamic_5_shot1

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: "This research aims to give a new insight to the relationship between host local immune response and the biological behaviour of the tumor by evaluating of intratumoral natural killer ( NK ) cells and tumor necrosis factor-alpha ( TNFalpha ) expressions in oral squamous cell carcinomas . New paraffin sections of the deepest parts of the 46 cases of oral squamous cell carcinomas were immunohistochemically treated by CD57 , selected as NK cell indicator , and TNFalpha monoclonal antibodies . The tumors were graded according to histopathologic grading scores of invasive margins and categorized into 2 groups as "" good "" and "" poor "" prognostic groups . Fifteen cases , from which could be obtained full clinical data , were clinically staged and because of the scarcity of the cases in each group were divided , again , two groups as group 1 : stage I+stage II and group 2 : stage III+stage IV . The expression levels of CD57 and TNFalpha were evaluated according to histopathologic grading groups and clinical staging groups . The results showed that the density of CD57+cells ( NK cells ) was statistically lower in tumors graded as poor prognostic group compared to the cases in good prognostic group . On the contrary , expression level of TNFalpha was statistically higher in poor prognostic group . These findings suggested that increased secretion of TNFalpha in the tumors , which show high invasive potential , may be one of the facilitating factors for tumor invasion and be responsible from suppression of NK cells . Withdrawal of NK cells in the high invasive tumor areas also reminds the necessity of certain shared genetic rearrangements in tumor cells for getting protected from NK cell attacks and moving ahead within the extracellular matrix ." OUTPUT: activating invasion and metastasis INPUT: "Here , we provide the necessary proof of concept , that it is possible to metabolically create a non-permissive or "" hostile "" stromal microenvironment , which actively prevents tumor engraftment in vivo . We developed a novel genetically engineered fibroblast cell line that completely prevents tumor formation in mice , with a 100% protection rate . No host side effects were apparent.This could represent a viable cellular strategy for preventing and treating a variety of human cancers . More specifically , we examined the autocrine and paracrine effects of the cellular delivery of TNF-alpha on breast cancer tumor growth and cancer metabolism . For this purpose , we recombinantly overexpressed TNF-alpha in human breast cancer cells ( MDA-MB-231 ) or human immortalized fibroblasts ( hTERT-BJ1 ) . Our results directly show that TNF-alpha functions as a potent tumor suppressor . Remarkably , TNF-alpha-expressing breast cancer cells were viable , without any significant increases in their basal apoptotic rate . However , after 4 weeks post-implantation , TNF-alpha-expressing breast cancer cells failed to form any tumors in xenografted mice ( 0 tumors/10 injections ) , ultimately conferring 100% protection against tumorigenesis . Similarly , TNF-alpha-overexpressing fibroblasts were also viable , without any increases in apoptosis . Significantly , complete tumor suppression was obtained by co-injecting TNF-alpha expressing stromal fibroblasts with human breast cancer cells , indicating that paracrine cell-mediated delivery of TNF-alpha can also prevent tumor engraftment and growth ( 0 tumors/10 injections ) . Mechanistically , TNF-alpha induced autophagy and mitochondrial dysfunction in both epithelial cancer cells and stromal fibroblasts , preventing energy transfer from the tumor microenvironment , likely "" starving "" the cancer cells to death . In addition , via qRT-PCR analysis of MDA-MB-231 cells , we observed that TNF-alpha mediated the up-regulation of gene transcripts associated with inflammation and senescence [ IL-1-beta , IL-6 , IL-8 , MCP-1 , COX-2 , p21(WAF1/CIP1) ] and downregulated known tumor-promoting genes ( collagen VI and MMP2 ) . Recombinant overexpression of TNF-alpha receptor(s) in MDA-MB-231 cells also significantly reduced tumor growth , but was not as effective as the TNF-alpha ligand itself in preventing tumor growth . Thus , we propose that stromal cell-mediated delivery of TNF-alpha to human tumors [ using transfected fibroblasts or mesenchymal stem cells ( hMSCs) ] may be a novel and effective strategy for the prevention and treatment of human cancers ." OUTPUT: resisting cell death;enabling replicative immortality;tumor promoting inflammation INPUT: "BACKGROUND The epidermal growth factor receptor ( EGFR ) is a validated therapeutic target in non-small cell lung cancer ( NSCLC ) . However , current single agent receptor targeting does not achieve a maximal therapeutic effect , and some mutations confer resistance to current available agents . In the current study we have examined , in different NSCLC cell lines , the combined effect of RNA interference targeting the EGFR mRNA , and inactivation of EGFR signaling using different receptor tyrosine kinase inhibitors ( TKIs ) or a monoclonal antibody cetuximab . METHODS NSCLC cells ( cell lines HCC827 , H292 , H358 , H1650 , and H1975 ) were transfected with EGFR siRNA and/or treated with the TKIs gefitinib , erlotinib , and afatinib , and/or with the monoclonal antibody cetuximab . The reduction of EGFR mRNA expression was measured by real-time quantitative RT-PCR . The down-regulation of EGFR protein expression was measured by western blot , and the proliferation , viability , caspase3/7 activity , and apoptotic morphology were monitored by spectrophotometry , fluorimetry , and fluorescence microscopy . The combined effect of EGFR siRNA and different drugs was evaluated using a combination index . RESULTS EGFR-specific siRNA strongly inhibited EGFR protein expression almost equally in all cell lines and inhibited cell growth and induced cell apoptosis in all NSCLC cell lines studied , albeit with a different magnitude . The effects on growth obtained with siRNA was strikingly different from the effects obtained with TKIs . The effects of siRNA probably correlate with the overall oncogenic significance of the receptor , which is only partly inhibited by the TKIs . The cells which showed weak response to TKIs , such as the H1975 cell line containing the T790M resistance mutation , were found to be responsive to siRNA knockdown of EGFR , as were cell lines with downstream TKI resistance mutations . The cell line HCC827 , harboring an exon 19 deletion mutation , was more than 10-fold more sensitive to TKI proliferation inhibition and apoptosis induction than any of the other cell lines . Cetuximab alone had no relevant in vitro activity at concentrations obtainable in the clinic . The addition of EGFR siRNA to either TKIs or cetuximab additively enhanced growth inhibition and induction of apoptosis in all five cell lines , independent of the EGFR mutation status ( wild-type or sensitizing mutation or resistant mutation ) . The strongest biological effect was observed when afatinib was combined with an EGFR-specific siRNA . CONCLUSIONS EGFR knockdown by siRNA further decreases the cell growth of lung cancer cells that are treated with TKIs or cetuximab alone , confirming that single agent drug targeting does not achieve a maximal biological effect . The siRNA inhibits EGFR oncogenic activity that bypasses downstream "" resistance "" mutations such as KRAS and PTEN . The combined treatment of siRNA and EGFR inhibitory agents is additive . The combination of a potent , irreversible kinase inhibitor such as afatinib , with EGFR-specific siRNAs should be further investigated as a new strategy in the treatment of lung cancer and other EGFR dependent cancers , including those with downstream resistance mutations ." OUTPUT: resisting cell death;sustaining proliferative signaling;genomic instability and mutation INPUT: "Intracellular pH ( pHi ) plays a critical role in the entry of cells into the DNA-synthesis phase of the cell cycle . Alterations in pHi may contribute to abnormal proliferative responses such as those seen in tumorigenic cells . We observed that alkaline stress leads to genomic transformation of Madin-Darby canine kidney ( MDCK ) cells . Transformed cells ( F cells ) form "" foci "" in culture , lack contact inhibition , and are able to migrate , typical characteristics of dedifferentiated tumorigenic cells . F cells exhibit spontaneous biorhythmicity . Rhythmic transmembrane Ca2+ flux activates plasma membrane K+ channels and Na+/H+ exchange . This leads to periodic changes of membrane voltage and pHi at about one cycle per minute . We conclude that endogenous oscillatory activity could be a trigger mechanism for DNA synthesis , proliferation , and abnormal growth of renal epithelial cells in culture ." OUTPUT: evading growth suppressors INPUT: "A distinct group of breast cancers , called "" basal "" or "" triple-negative "" ( TN ) cancers express both basal cytokeratins and the epidermal growth factor receptor , but fail to express estrogen receptors , progesterone receptors or HER2 and have stem-like or mesenchymal features . They are particularly aggressive , are frequently chemo-resistant , with p53 mutation , up-regulation of IL-6 and Stat3 . Because TN cells are particularly sensitive to the anti-diabetic agent metformin , we hypothesized that it may target JAK2/Stat3 signaling . The effects of metformin upon Stat3 expression and activation were examined in four human TN cell lines . Metformin's effects were also studied in sublines with forced over-expression of constitutively active ( CA ) Stat3 , as well as lines with stable knockdown of Stat3 . Metformin inhibited Stat3 activation ( P-Stat3 ) at Tyr705 and Ser727 and downstream signaling in each of the four parental cell lines . CA-Stat3 transfection attenuated , whereas Stat3 knockdown enhanced , the effects of metformin upon growth inhibition and apoptosis induction . A Stat3 specific inhibitor acted synergistically with metformin in reducing cell growth and inducing apoptosis . An mTOR inhibitor showed no significant interaction with metformin . In summary , Stat3 is a critical regulator of metformin action in TN cancer cells , providing the potential for enhancing metformin's efficacy in the clinical setting ." OUTPUT: resisting cell death INPUT: "We have used a combination of vitamin A ( all-trans-retinyl palmitate ) , 5-fluorouracil ( 5-FU ) and radiation to treat human head and neck squamous cell carcinoma ( HNSCC ) . This chemoradiotherapy is called "" FAR therapy. "" In this study we examined the effects of all-trans-retinoic acid ( ATRA ) , the active metabolite of vitamin A , and ATRA plus 5-FU on two HNSCC cell lines ( YCU-N861 and YCU-H891 ) to gain insight into the molecular mechanisms of FAR therapy . ATRA at 1 mM ( the order of concentration found in HNSCC tumors treated with FAR therapy ) inhibited cell proliferation and caused G1 cell cycle arrest in both cell lines . This was associated with a decrease in cyclin D1 , an increase in p27(Kip1) and a reduction in the hyperphosphorylated form of retinoblastoma protein ( pRB ) . With YCU-N861 cells , ATRA also caused a decrease in Bcl-2 and Bcl-X(L) and an increase in Bax . Both ATRA and 5-FU activated c-Jun N-terminal kinase ( JNK ) 1 and the combination of both agents resulted in additive or synergistic activation of JNK1 , and also enhanced the induction of apoptosis . The YCU-H891 cells , in which the epidermal growth factor receptor ( EGFR)-signal transducer and activator of transcription 3 ( Stat3 ) pathway is constitutively activated , were more resistant to treatments with ATRA , 5-FU and the combination of both agents than YCU-N861 cells . A dominant negative Stat3 construct strongly enhanced the cellular sensitivity of this cell line to 5-FU but not to ATRA . In addition there is evidence that activation of Stat3 is associated with cellular resistance to radiation in HNSCC . Therefore , the addition to FAR therapy of agents that inhibit activation of the Stat3 pathway may enhance the clinical response of patients with HNSCC to FAR therapy ." OUTPUT:

sustaining proliferative signaling;evading growth suppressors;resisting cell death

HoC_dynamic_5_shot2

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND : Previously , we have observed that highly unsaturated dietary ( n-3 ) fatty acids inhibit cell proliferation in conjunction with stimulation of insulin-like growth factor-binding protein ( IGFBP)-6 secretion in Caco-2 cells , a human colon carcinoma cell line . METHODS : To test the converse hypothesis that inhibition of endogenous IGFBP-6 secretion stimulates Caco-2 cell proliferation , cells were transfected with the antisense IGFBP-6 expression construct or pcDNA3 vector only , and single colonies resistant to G418 sulfate were isolated . RESULTS : Our initial studies indicated that three antisense clones grew faster and produced less IGFBP-6 than two pcDNA3 clones , so antisense IGFBP-6 #5 and pcDNA3 #8 were selected for further detailed analysis . Both the control and antisense clones grew in serum-free medium reaching a plateau density at day eight . However , the antisense clone grew at a rate faster than that of the control and reached a final density that was 31 +/- 3% higher than the control . Northern blot , ligand blot and immunoblot analyses revealed that accumulation of IGFBP-6 mRNA and concentrations of IGFBP-6 peptide produced by the antisense clone were decreased by 80-90% compared to the control . The doubling times of the antisense and control clones were 21.9 +/- 0.4 and 24.8 +/- 0.3 h ( P &lt ; 0.05 ) , respectively . Exogenous IGF-I and IGF-II ( 0.2-200 nmol/L ) stimulated proliferation of both the control and antisense clones in a dose-dependent manner , but the relative potency and efficacy of IGF-II was higher in the antisense clone compared to the control . These results indicate that suppression of IGFBP-6 secretion correlates with an increase in the basal rate of Caco-2 cell growth . CONCLUSIONS : Our findings are consistent with the hypothesis that IGFBP-6 inhibits cell growth by binding to endogenously produced IGF-II , thereby preventing IGF-II from interacting with the IGF-I receptor to stimulate cellular proliferation by an autocrine mechanism . OUTPUT: sustaining proliferative signaling INPUT: The insulin-like growth factor ( IGF ) pathway represents one of the most studied molecular regulatory networks in oncology . Clinical trials investigating the therapeutic value of anti-IGF1 receptor ( IGF1R ) therapies in cancer , including prostate cancer , are ongoing . However , the multiple functions of the IGF network in the prostate are not entirely known . To elucidate the effects of IGF and insulin ( INS ) on prostate cells , we stimulated prostate cancer ( PC3 , DU145 , LNCaP , DUCaP ) and noncancerous prostate cells ( EP156T , RWPE-1 ) and observed differing responses : whereas cancer cells responded to IGF and INS exposure by way of enhanced cell proliferation and glucose consumption , basal to luminal differentiation was induced in noncancerous cells . The same diverse responses were observed when the growth factor receptors IGF1R or INSR were overexpressed . Down-regulation of IGF1R or INSR isoform A ( INSRA ) also inhibited only proliferation of cancer cells . The proliferative response induced by the INSR in cancer cells was mediated solely by the INSRA . Moreover we observed that the receptors of the IGF network mutually influence their expression and exert redundant functions , thus underscoring the functional molecular network formed by IGF , INS , IGF1R , and INSR . Collectively we found that both IGF1R and INSRA have oncogenic effects in prostate cancer , but the IGF network also has important physiological functions in the noncancerous prostate . These data provide new insights into the biology of the IGF network in the prostate , thereby facilitating the design and interpretation of clinical studies investigating IGF1R targeting agents . OUTPUT: sustaining proliferative signaling INPUT: Insulin-like growth factor ( IGF)-I receptor ( IGF-IR ) signaling is required for carcinogenicity and progression of several cancers but the function of this pathway and its utility as a therapeutic target have not been studied comprehensively in biliary tract carcinomas ( BTC ) . We investigated the immunohistochemical expression of elements of the IGF axis , matrilysin , overexpression of p53 and the methylation status of the IGFBP-3 promoter in 80 surgically resected BTC . We also assessed the effect of IGF-IR blockade on signal transduction , proliferation and survival in three BTC cell lines using a new tyrosine kinase inhibitor , BMS-536924 , and dominant negative IGF-IR ( IGF-IR/dn ) . The effects of IGF-IR blockade was also studied in nude mouse xenograft models . IGF-I was expressed in 60% and IGF-II in 50% of tumors . High expression was associated with tumor size . IGF-IR was expressed in 69% of the cases and was associated with advanced stage and matrilysin expression . Hypermethylation of the IGFBP-3 promoter was detected in 41% of BTC and was inversely correlated with p53 expression . BMS-536924 blocked autophosphorylation of IGF-IR and both Akt and ERK activation by both IGF-I and insulin . BMS-536924 suppressed proliferation and tumorigenicity in vitro in a dose-dependent fashion . This inhibitor upregulated chemotherapy-induced apoptosis in a dose-dependent fashion . Moreover , IGF-IR blockade was effective against tumors in mice . IGF-IR might identify a subset of BTC with a particularly aggressive phenotype and is a candidate therapeutic target in this disease . BMS-536924 might have significant therapeutic utility . OUTPUT: resisting cell death INPUT: OBJECTIVES Epidermal growth factor ( EGF ) stimulates cell proliferation by binding to its receptor ( epidermal growth factor receptor ) , and the overexpression of this receptor is associated with poorer prognosis . The EGF gene presents a polymorphism at position 61 ( A/G ) , associated with higher EGF production . We examined the association between this polymorphism and cervical cancer through a case-control study . METHODS This study used the PCR-restriction fragment length polymorphism method on a sample of 384 women with cervical lesions and 500 controls of white ethnicity . RESULTS In cases of cervical cancer , we found an increased risk of progression to advanced disease ( The International Federation of Gynecology and Obstetrics stage IIb/IV ) ( odds ratios=2.05 ; 95% confidence intervals=1.11 to 3.79 ; P=0.021 ) , and this risk was particularly evident in G carriers for younger women ( odds ratios=2.96 ; 95% confidence intervals=1.41-6.20 , P=0.003 ) . CONCLUSIONS We hypothesize the onset of an advanced disease-driven selective pressure due to the effect of oncogenic human papillomavirus types in a favorable genetic background observed in G carrier women . These results suggest that EGF functional polymorphism may influence cervical cancer prognosis through an EGF/epidermal growth factor receptor pathway . OUTPUT: sustaining proliferative signaling INPUT: Previously we have found deregulation of collagen metabolism in human pancreatitis and pancreatic cancer tissues . Insulin-like growth factor-I ( IGF-I ) is known to stimulate collagen biosynthesis through interaction with IGF-I receptor . IGF-I binding proteins ( BPs ) regulate the activity of IGF-I . We investigated whether serum and tissue IGF-I and IGF-BPs as well as tissue IGF-I receptor expression may reflect disturbances of collagen metabolism in patients with pancreatitis and pancreatic cancer . In pancreatitis tissue , a significant increase in IGF-I and IGFBP-3 content was accompanied by a distinct increase in IGF-I receptor expression , compared to control pancreas tissue . In contrast , serum from patients with pancreatitis did not show significant increases in IGF-I and IGFBP-3 levels , however , significant increases in IGFBP-1 level ( 2.5 fold ) . Moreover , a distinct decrease in radioactive IGF-binding to the BPs , compared to control serum , was found . Pancreatic cancer tissue and serum of patients with pancreatic cancer showed significant increases in IGF-I , IGFBP-3 and IGFBP-1 content , accompanied by dramatic increases in IGF-I tissue receptor expression , compared to controls . In serum of patients with pancreatic cancer distinct increases in radioactive IGF-binding to 46 kDa BP , compared to control serum , were observed . The data suggest that disturbances in tissue collagen metabolism during pancreatic diseases may result from deregulation of IGF-I homeostasis and that elevated serum levels of IGF-I , IGFBP-3 and IGFBP-1 may serve as markers of pancreatic cancer . OUTPUT: sustaining proliferative signaling INPUT: BACKGROUND Insulin-like growth factor I ( IGF-I ) stimulates cell proliferation and inhibits apoptosis in the lung and other tissues by interacting with the IGF-I receptor . The major binding protein for IGF-I , insulin-like growth factor-binding protein 3 ( IGFBP-3 ) , modulates the effects of IGF-I but also inhibits cell growth and induces apoptosis independent of IGF-I and its receptor . In a prospective study of men in Shanghai , China , we examined the association between serum levels of IGF-I and IGFBP-3 and the subsequent risk of lung cancer . METHODS From 1986 to 1989 , serum was collected from 18,244 men aged 45-64 years living in Shanghai without a history of cancer . We analyzed IGF-I and IGFBP-3 levels in serum from 230 case patients who developed incident lung cancer during follow-up and from 740 control subjects . RESULTS Among 230 case patients and 659 matched control subjects , increased IGF-I levels were not associated with increased risk of lung cancer . However , for subjects in the highest quartile relative to the lowest quartile of IGFBP-3 , the odds ratio ( OR ) for lung cancer , adjusted for smoking and IGF-I , was 0.50 ( 95% confidence interval [ CI ] = 0.25 to 1.02 ) . When the analysis was restricted to ever smokers ( 184 case patients and 344 matched control subjects ) , the OR for lung cancer in men in the highest quartile of IGFBP-3 relative to those in the lowest quartile , adjusted for smoking and IGF-I , was 0.41 ( 95% CI = 0.18 to 0.92 ) . CONCLUSIONS In this prospective study of Chinese men , higher serum levels of IGF-I did not increase the risk of lung cancer . However , subjects with higher serum levels of IGFBP-3 were at reduced risk of lung cancer . This finding is consistent with experimental data that indicate that IGFBP-3 can inhibit cellular proliferation and induce apoptosis independent of IGF-I and the IGF-I receptor . OUTPUT:

sustaining proliferative signaling;resisting cell death

HoC_dynamic_5_shot3

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Often the use of cytotoxic drugs in cancer therapy results in stable disease rather than regression of the tumor , and this is typically seen as a failure of treatment . We now show that DNA damage is able to induce senescence in tumor cells expressing wild-type p53 . We also show that cytotoxics are capable of inducing senescence in tumor tissue in vivo . Our results suggest that p53 and p21 play a central role in the onset of senescence , whereas p16(INK4a) function may be involved in maintaining senescence . Thus , like apoptosis , senescence appears to be a p53-induced cellular response to DNA damage and an important factor in determining treatment outcome . OUTPUT: enabling replicative immortality;genomic instability and mutation;resisting cell death INPUT: When the prostate cancer cells become unresponsive to androgen therapy , resistance to chemotherapy becomes imminent , resulting in high mortality . To combat this situation , lycopodine , a pharmacologically important bioactive component derived from Lycopodium clavatum spores , was tested against hormone sensitive ( LnCaP ) and refractory ( PC3 ) prostate cancer cells in vitro . This study aims to check if lycopodine has demonstrable anti-cancer effects and if it has , to find out the possible mechanism of its action . The MTT assay was performed to evaluate the cytotoxic effect . Depolarization of mitochondrial membrane potential , cell cycle , EGF receptor activity and apoptosis were recorded by FACS ; profiles of different anti- and pro-apoptotic genes and their products were studied by semi-quantitative RT-PCR , indirect-ELISA , western blotting . Drug-DNA interaction was determined by CD spectroscopy . Administration of lycopodine down-regulated the expression of 5-lipoxygenase and the 5-oxo-ETE receptor ( OXE receptor1 ) and EGF receptor , and caused up-regulation of cytochrome c with depolarization of mitochondrial inner membrane potential , without palpable change in p53 activity , resulting in apoptosis , cell arrest at G0/G1 stage and ultimately reduced proliferation of cancer cells ; concomitantly , there was externalization of phosphotidyl serine residues . CD spectroscopic analysis revealed intercalating property of lycopodine with DNA molecule , implicating its ability to block cellular DNA synthesis . The overall results suggest that lycopodine is a promising candidate suitable for therapeutic use as an anti-cancer drug . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: DNA polymerases beta ( pol beta ) and eta ( pol eta ) are the only two eukaryotic polymerases known to efficiently bypass cisplatin and oxaliplatin adducts in vitro . Frameshift errors are an important aspect of mutagenesis . We have compared the types of frameshifts that occur during translesion synthesis past cisplatin and oxaliplatin adducts in vitro by pol beta and pol eta on a template containing multiple runs of nucleotides flanking a single platinum-GG adduct . Translesion synthesis past platinum adducts by pol beta resulted in approximately 50% replication products containing single-base deletions . For both adducts the majority of -1 frameshifts occurred in a TTT sequence 3-5 bp upstream of the DNA lesion . For pol eta , all of the bypass products for both cisplatin and oxaliplatin adducts contained -1 frameshifts in the upstream TTT sequence and most of the products of replication on oxaliplatin-damaged templates had multiple replication errors , both frameshifts and misinsertions . In addition , on platinated templates both polymerases generated replication products 4-8 bp shorter than the full-length products . The majority of short cisplatin-induced products contained an internal deletion which included the adduct . In contrast , the majority of oxaliplatin-induced short products contained a 3 ' terminal deletion . The implications of these in vitro results for in vivo mutagenesis are discussed . OUTPUT: genomic instability and mutation INPUT: Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers . A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation . We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products . This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine ( NMBzA ) , and the response that increased levels of dietary vitamin E has on this induced carcinogenesis . Female C57BL/6 mice received NMBzA or vehicle ( corn oil ) i.p. weekly for 3 weeks . Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet . As an assessment of free radical activity , exhaled ethane was measured prior to killing the animals at 26 weeks . Esophagi from the various mice groups were assessed for size and frequency of tumors . Livers homogenates were analyzed for vitamins A and E , lipid fluorescence , conjugated dienes and malondialdehyde . Hepatic levels of vitamin A and E were decreased ( P &lt ; 0.05 ) and indices of lipid peroxidation were greater ( P &lt ; 0.05 ) in NMBzA-treated mice relative to controls . Lipid peroxidation and serum transaminases ( ALT and AST ) were greatest in mice given NMBzA and infected with the retroviruses . Incidence of esophageal tumors were also greatest in the NMBzA-treated , immunocompromised animals . Mice fed vitamin E-supplemented diets showed increased ( P &lt ; 0.05 ) hepatic concentrations of vitamin E and vitamin A , decreased activities of serum transaminases , decreased indices of lipid peroxidation , and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice . These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals . OUTPUT: avoiding immune destruction INPUT: The INK4a/ARF tumor suppressor locus is implicated in the senescence-like growth arrest provoked by oncogenic Ras in primary cells . INK4a and ARF are distinct proteins encoded by transcripts in which a shared exon is decoded in alternative reading frames . Here we analyze dermal fibroblasts ( designated Q34 ) from an individual carrying independent missense mutations in each copy of the common exon . Both mutations alter the amino acid sequence of INK4a and functionally impair the protein , although they do so to different degrees . Only one of the mutations affects the sequence of ARF , causing an apparently innocuous change near its carboxy terminus . Unlike normal human fibroblasts , Q34 cells are not permanently arrested by Ras or its downstream effectors Ets1 and Ets2 . Moreover , ectopic Ras enables the cells to grow as anchorage-independent colonies , and in relatively young Q34 cells anchorage independence can be achieved without addition of telomerase or perturbation of the p53 pathway . Whereas ARF plays the principal role in Ras-induced arrest of mouse fibroblasts , our data imply that INK4a assumes this role in human fibroblasts . OUTPUT: genomic instability and mutation INPUT: p53 and INK4a/ARF mutations promote tumorigenesis and drug resistance , in part , by disabling apoptosis . We show that primary murine lymphomas also respond to chemotherapy by engaging a senescence program controlled by p53 and p16(INK4a) . Hence , tumors with p53 or INK4a/ARF mutations-but not those lacking ARF alone-respond poorly to cyclophosphamide therapy in vivo . Moreover , tumors harboring a Bcl2-mediated apoptotic block undergo a drug-induced cytostasis involving the accumulation of p53 , p16(INK4a) , and senescence markers , and typically acquire p53 or INK4a mutations upon progression to a terminal stage . Finally , mice bearing tumors capable of drug-induced senescence have a much better prognosis following chemotherapy than those harboring tumors with senescence defects . Therefore , cellular senescence contributes to treatment outcome in vivo . OUTPUT:

enabling replicative immortality;genomic instability and mutation;resisting cell death

HoC_dynamic_5_shot4

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Transforming growth factor beta ( TGF-β ) signaling has been implicated in driving tumor progression and metastasis by inducing stem cell-like features in some human cancer cell lines . In this study , we have utilized a novel murine cell line NMuMG-ST , which acquired cancer stem cell ( CSC ) phenotypes during spontaneous transformation of the untransformed murine mammary cell line NMuMG , to investigate the role of autocrine TGF-β signaling in regulating their survival , metastatic ability , and the maintenance of cancer stem cell characteristics . We have retrovirally transduced a dominant-negative TGF-β type II receptor ( DNRII ) into the NMuMG-ST cell to abrogate autocrine TGF-β signaling . The expression of DNRII reduced TGF-β sensitivity of the NMuMG-ST cells in various cell-based assays . The blockade of autocrine TGF-β signaling reduced the ability of the cell to grow anchorage-independently and to resist serum deprivation-induced apoptosis . These phenotypes were associated with reduced levels of active and phosphorylated AKT and ERK , and Gli1 expression suggesting that these pathways contribute to the growth and survival of this model system . More interestingly , the abrogation of autocrine TGF-β signaling also led to the attenuation of several features associated with mammary stem cells including epithelial-mesenchymal transition , mammosphere formation , and expression of stem cell markers . When xenografted in athymic nude mice , the DNRII cells were also found to undergo apoptosis and induced significantly lower lung metastasis burden than the control cells even though they formed similar size of xenograft tumors . Thus , our results indicate that autocrine TGF-β signaling is involved in the maintenance and survival of stem-like cell population resulting in the enhanced metastatic ability of the murine breast cancer cells . OUTPUT: resisting cell death;activating invasion and metastasis INPUT: Annexin II is secreted into the extracellular environment , where , via interactions with specific proteases and extracellular matrix proteins , it participates in plasminogen activation , cell adhesion , and tumor metastasis and invasion . However , mechanisms regulating annexin II transport across the cellular membrane are unknown . In this study , we used coimmunoprecipitation to show that Annexin-II was bound to insulin and insulin-like growth factor-1 ( IGF-1 ) receptors in PC12 cells and NIH-3T3 cells overexpressing insulin ( NIH-3T3(IR) ) or IGF-1 receptor ( NIH-3T3(IGF-1R) ) . Stimulation of insulin and IGF-1 receptors by insulin caused a temporary dissociation of annexin II from these receptors , which was accompanied by an increased amount of extracellular annexin II detected in the media of PC12 , NIH-3T3(IR) , and NIH-3T3(IGF-1R) cells but not in that of untransfected NIH-3T3 cells . Activation of a different growth factor receptor , the platelet-derived growth factor receptor , did not produce such results . Tyrphostin AG1024 , a tyrosine kinase inhibitor of insulin and IGF-1 receptor , was shown to inhibit annexin II secretion along with reduced receptor phosphorylation . Inhibitors of a few downstream signaling enzymes including phosphatidylinositol 3-kinase , pp60c-Src , and protein kinase C had no effect on insulin-induced annexin II secretion , suggesting a possible direct link between receptor activation and annexin II secretion . Immunocytochemistry revealed that insulin also induced transport of the membrane-bound form of annexin II to the outside layer of the cell membrane and appeared to promote cell aggregation . These results suggest that the insulin receptor and its signaling pathways may participate in molecular mechanisms mediating annexin II secretion . OUTPUT: sustaining proliferative signaling INPUT: The insulin-like growth factor ( IGF ) pathway represents one of the most studied molecular regulatory networks in oncology . Clinical trials investigating the therapeutic value of anti-IGF1 receptor ( IGF1R ) therapies in cancer , including prostate cancer , are ongoing . However , the multiple functions of the IGF network in the prostate are not entirely known . To elucidate the effects of IGF and insulin ( INS ) on prostate cells , we stimulated prostate cancer ( PC3 , DU145 , LNCaP , DUCaP ) and noncancerous prostate cells ( EP156T , RWPE-1 ) and observed differing responses : whereas cancer cells responded to IGF and INS exposure by way of enhanced cell proliferation and glucose consumption , basal to luminal differentiation was induced in noncancerous cells . The same diverse responses were observed when the growth factor receptors IGF1R or INSR were overexpressed . Down-regulation of IGF1R or INSR isoform A ( INSRA ) also inhibited only proliferation of cancer cells . The proliferative response induced by the INSR in cancer cells was mediated solely by the INSRA . Moreover we observed that the receptors of the IGF network mutually influence their expression and exert redundant functions , thus underscoring the functional molecular network formed by IGF , INS , IGF1R , and INSR . Collectively we found that both IGF1R and INSRA have oncogenic effects in prostate cancer , but the IGF network also has important physiological functions in the noncancerous prostate . These data provide new insights into the biology of the IGF network in the prostate , thereby facilitating the design and interpretation of clinical studies investigating IGF1R targeting agents . OUTPUT: sustaining proliferative signaling INPUT: BACKGROUND : Previously , we have observed that highly unsaturated dietary ( n-3 ) fatty acids inhibit cell proliferation in conjunction with stimulation of insulin-like growth factor-binding protein ( IGFBP)-6 secretion in Caco-2 cells , a human colon carcinoma cell line . METHODS : To test the converse hypothesis that inhibition of endogenous IGFBP-6 secretion stimulates Caco-2 cell proliferation , cells were transfected with the antisense IGFBP-6 expression construct or pcDNA3 vector only , and single colonies resistant to G418 sulfate were isolated . RESULTS : Our initial studies indicated that three antisense clones grew faster and produced less IGFBP-6 than two pcDNA3 clones , so antisense IGFBP-6 #5 and pcDNA3 #8 were selected for further detailed analysis . Both the control and antisense clones grew in serum-free medium reaching a plateau density at day eight . However , the antisense clone grew at a rate faster than that of the control and reached a final density that was 31 +/- 3% higher than the control . Northern blot , ligand blot and immunoblot analyses revealed that accumulation of IGFBP-6 mRNA and concentrations of IGFBP-6 peptide produced by the antisense clone were decreased by 80-90% compared to the control . The doubling times of the antisense and control clones were 21.9 +/- 0.4 and 24.8 +/- 0.3 h ( P &lt ; 0.05 ) , respectively . Exogenous IGF-I and IGF-II ( 0.2-200 nmol/L ) stimulated proliferation of both the control and antisense clones in a dose-dependent manner , but the relative potency and efficacy of IGF-II was higher in the antisense clone compared to the control . These results indicate that suppression of IGFBP-6 secretion correlates with an increase in the basal rate of Caco-2 cell growth . CONCLUSIONS : Our findings are consistent with the hypothesis that IGFBP-6 inhibits cell growth by binding to endogenously produced IGF-II , thereby preventing IGF-II from interacting with the IGF-I receptor to stimulate cellular proliferation by an autocrine mechanism . OUTPUT: sustaining proliferative signaling INPUT: INTRODUCTION The insulin-like growth factor I receptor ( IGF-IR ) pathway plays a major role in cancer growth , tumor cell survival and resistance to therapy . BACKGROUND Preclinical evidence that targeting the IGF-IR is effective in cancer treatment has been accumulating for almost 2 decades . Early clinical trials revealed an acceptable safety profile together with pharmacodynamic evidence that the receptor can be targeted successfully . It is premature to draw conclusions regarding the therapeutic potential of this class of compounds but well-documented single-agent activity was noted during phase I evaluations , and recent evidence from a phase-II study suggests that co-administration of an anti-IGF-1R antibody with chemotherapy for non-small-cell lung cancer ( NSCLC ) improves objective response rate and progression-free survival . VIEWPOINTS These early results are a strong indication for continued research on the targeting of IGF-R , particularly in the treatment of NSCLC . CONCLUSIONS Today , IGF-1R targeting appears a promising approach , more than two dozen compounds have been developed and clinical trials are underway . OUTPUT: sustaining proliferative signaling INPUT: Insulin-like growth factor I ( IGF-I ) and IGF-II stimulate cancer cell proliferation via interaction with the type I IGF receptor ( IGF-IR ) . We put forward the hypothesis that IGF-IR mediates cancer cell growth by regulating amino acid transport , both when sufficient nutrients are present and when key nutrients such as glutamine are in limited supply . We examined the effects of alphaIR3 , the monoclonal antibody recognizing IGF-IR , on cell growth and amino acid transport across the cell membrane in a human neuroblastoma cell line , SK-N-SH . In the presence of alphaIR3 ( 2 micro/ml ) , cell proliferation was significantly attenuated in both control ( 2 mM glutamine ) and glutamine-deprived ( 0 mM glutamine ) groups . Glutamine deprivation resulted in significantly increased glutamate ( system X(AG)(-) ) , MeAIB ( system A ) , and leucine ( system L ) transport , which was blocked by alphaIR3 . Glutamine ( system ASC ) and MeAIB transport was significantly decreased by alphaIR3 in the control group . Addition of alphaIR3 significantly decreased DNA and protein biosynthesis in both groups . Glutamine deprivation increased the IGF-IR protein on the cell surface . Our results suggest that activation of IGF-IR promotes neuroblastoma cell proliferation by regulating trans-membrane amino acid transport . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot5

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: We found that the human colon cancer cell line SW480 consists of two distinct subpopulations which we have designated E-type ( epithelial ) and R-type ( round ) . Pure cultures of each type were obtained by subcloning , and both have maintained their characteristic phenotypes for at least 1 year ( 40 passages ) . E-type cells are the major ( &gt ; 98% ) type in the parental SW480 cell line . They form flat epithelial-like colonies . In contrast , R-type cells , which constitute a minor fraction ( &lt ; 2% ) of the parental cell line , have a rounded shape and grow in clusters of piled-up cells . Compared to E-type cells or the parental SW480 cells , isolated R-type cells display decreased doubling time , loss of contact inhibition , less adhesiveness to culture plates , higher anchorage-independent growth in soft agar , and a much more aneuploid karyotype . When injected s.c. into nude mice , R-type cells produce much larger tumors within the same period of time than E-type cells , and the tumors are less differentiated than those produced by the E-type cells . Cell fusion experiments between R-type and E-type cells revealed that the R-type phenotype is dominant , and the results suggest that this is due to one or a few genetic changes . Taken together , these findings suggest that the R-type cells represent a more malignant variant of the E-type cells . They may be useful , therefore , for studying mechanisms involved in tumor progression . OUTPUT: evading growth suppressors INPUT: The immune system of vertebrates is able to detect bacterial DNA based on the presence of unmethylated CpG motifs . We examined the therapeutic potential of oligodeoxynucleotides with CpG motifs ( CpG ODN ) in a colon carcinoma model in BALB/c mice . Tumors were induced by s.c. injection of syngeneic C26 cells or Renca kidney cancer cells as a control . Injection of CpG ODN alone or in combination with irradiated tumor cells did not protect mice against subsequent tumor challenge . In contrast , weekly injections of CpG ODN into the margin of already established tumors resulted in regression of tumors and complete cure of mice . The injection site was critical , since injection of CpG ODN at distant sites was not effective . Mice with two bilateral C26 tumors rejected both tumors upon peritumoral injection of one tumor , indicating the development of a systemic immune response . The tumor specificity of the immune response was demonstrated in mice bearing a C26 tumor and a Renca tumor at the same time . Mice that rejected a tumor upon peritumoral CpG treatment remained tumor free and were protected against rechallenge with the same tumor cells , but not with the other tumor , demonstrating long term memory . Tumor-specific CD8 T cells as well as innate effector cells contributed to the antitumor activity of treatment . In conclusion , peritumoral CpG ODN monotherapy elicits a strong CD8 T cell response and innate effector mechanisms that seem to act in concert to overcome unresponsiveness of the immune system toward a growing tumor . OUTPUT: avoiding immune destruction INPUT: Transforming growth factor beta ( TGF-β ) signaling has been implicated in driving tumor progression and metastasis by inducing stem cell-like features in some human cancer cell lines . In this study , we have utilized a novel murine cell line NMuMG-ST , which acquired cancer stem cell ( CSC ) phenotypes during spontaneous transformation of the untransformed murine mammary cell line NMuMG , to investigate the role of autocrine TGF-β signaling in regulating their survival , metastatic ability , and the maintenance of cancer stem cell characteristics . We have retrovirally transduced a dominant-negative TGF-β type II receptor ( DNRII ) into the NMuMG-ST cell to abrogate autocrine TGF-β signaling . The expression of DNRII reduced TGF-β sensitivity of the NMuMG-ST cells in various cell-based assays . The blockade of autocrine TGF-β signaling reduced the ability of the cell to grow anchorage-independently and to resist serum deprivation-induced apoptosis . These phenotypes were associated with reduced levels of active and phosphorylated AKT and ERK , and Gli1 expression suggesting that these pathways contribute to the growth and survival of this model system . More interestingly , the abrogation of autocrine TGF-β signaling also led to the attenuation of several features associated with mammary stem cells including epithelial-mesenchymal transition , mammosphere formation , and expression of stem cell markers . When xenografted in athymic nude mice , the DNRII cells were also found to undergo apoptosis and induced significantly lower lung metastasis burden than the control cells even though they formed similar size of xenograft tumors . Thus , our results indicate that autocrine TGF-β signaling is involved in the maintenance and survival of stem-like cell population resulting in the enhanced metastatic ability of the murine breast cancer cells . OUTPUT: resisting cell death;activating invasion and metastasis INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: In vitro model systems for studying uterine leiomyomas are limited in that human-derived leiomyoma cells grow poorly in culture compared with normal myometrial cells and begin to senesce early , at approximately passage 10 in our studies . To our knowledge , a good in vitro human-derived cell culturing system for leiomyomas does not exist . In an attempt to fill this void , we have immortalized a uterine leiomyoma cell line by inducing telomerase activity , which allows cells to bypass their normal programmed senescence . Telomerase activity was induced by infecting the target ( uterine leiomyoma and normal myometrial ) cells with a retroviral vector containing hTERT , the gene for the catalytic subunit of telomerase . Subsequent analysis by RT-PCR and the telomeric repeat amplification protocol assay confirmed expression of the inserted gene and induction of telomerase activity in leiomyoma and myometrial cells . Analysis of cells for estrogen receptor-alpha and progesterone receptor proteins by Western blotting showed no change in expression of these proteins between the immortalized and parental leiomyoma and myometrial cells . Both immortalized and parental myometrial and leiomyoma cells expressed the smooth muscle-specific cytoskeletal protein alpha-actin and were negative for mutant p53 protein as evidenced by immunocytochemical staining . The immortalized leiomyoma and myometrial cells showed no anchorage-independent growth , with the exception of a small subpopulation of immortalized leiomyoma cells at a higher passage that did form two to three small colonies ( per 50,000 cells ) in soft agar . None of the immortalized cells were tumorigenic in nude mice . In conclusion , our data show the successful insertion of the hTERT gene into leiomyoma and myometrial cells and the immortalization of these cell lines without phenotypic alteration from the parental cell types ( up to 200 population doublings ) . These cells should help to advance research in understanding the molecular pathways involved in the conversion of a normal myometrial cell to a leiomyoma cell and the mechanisms responsible for the growth of uterine leiomyomas . Answers to these questions will undoubtedly lead to the development of more effective treatment and intervention regimens for clinical cases of uterine leiomyoma . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot6

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: "BACKGROUND The epidermal growth factor receptor ( EGFR ) is a validated therapeutic target in non-small cell lung cancer ( NSCLC ) . However , current single agent receptor targeting does not achieve a maximal therapeutic effect , and some mutations confer resistance to current available agents . In the current study we have examined , in different NSCLC cell lines , the combined effect of RNA interference targeting the EGFR mRNA , and inactivation of EGFR signaling using different receptor tyrosine kinase inhibitors ( TKIs ) or a monoclonal antibody cetuximab . METHODS NSCLC cells ( cell lines HCC827 , H292 , H358 , H1650 , and H1975 ) were transfected with EGFR siRNA and/or treated with the TKIs gefitinib , erlotinib , and afatinib , and/or with the monoclonal antibody cetuximab . The reduction of EGFR mRNA expression was measured by real-time quantitative RT-PCR . The down-regulation of EGFR protein expression was measured by western blot , and the proliferation , viability , caspase3/7 activity , and apoptotic morphology were monitored by spectrophotometry , fluorimetry , and fluorescence microscopy . The combined effect of EGFR siRNA and different drugs was evaluated using a combination index . RESULTS EGFR-specific siRNA strongly inhibited EGFR protein expression almost equally in all cell lines and inhibited cell growth and induced cell apoptosis in all NSCLC cell lines studied , albeit with a different magnitude . The effects on growth obtained with siRNA was strikingly different from the effects obtained with TKIs . The effects of siRNA probably correlate with the overall oncogenic significance of the receptor , which is only partly inhibited by the TKIs . The cells which showed weak response to TKIs , such as the H1975 cell line containing the T790M resistance mutation , were found to be responsive to siRNA knockdown of EGFR , as were cell lines with downstream TKI resistance mutations . The cell line HCC827 , harboring an exon 19 deletion mutation , was more than 10-fold more sensitive to TKI proliferation inhibition and apoptosis induction than any of the other cell lines . Cetuximab alone had no relevant in vitro activity at concentrations obtainable in the clinic . The addition of EGFR siRNA to either TKIs or cetuximab additively enhanced growth inhibition and induction of apoptosis in all five cell lines , independent of the EGFR mutation status ( wild-type or sensitizing mutation or resistant mutation ) . The strongest biological effect was observed when afatinib was combined with an EGFR-specific siRNA . CONCLUSIONS EGFR knockdown by siRNA further decreases the cell growth of lung cancer cells that are treated with TKIs or cetuximab alone , confirming that single agent drug targeting does not achieve a maximal biological effect . The siRNA inhibits EGFR oncogenic activity that bypasses downstream "" resistance "" mutations such as KRAS and PTEN . The combined treatment of siRNA and EGFR inhibitory agents is additive . The combination of a potent , irreversible kinase inhibitor such as afatinib , with EGFR-specific siRNAs should be further investigated as a new strategy in the treatment of lung cancer and other EGFR dependent cancers , including those with downstream resistance mutations ." OUTPUT: resisting cell death;sustaining proliferative signaling;genomic instability and mutation INPUT: Annexin II is secreted into the extracellular environment , where , via interactions with specific proteases and extracellular matrix proteins , it participates in plasminogen activation , cell adhesion , and tumor metastasis and invasion . However , mechanisms regulating annexin II transport across the cellular membrane are unknown . In this study , we used coimmunoprecipitation to show that Annexin-II was bound to insulin and insulin-like growth factor-1 ( IGF-1 ) receptors in PC12 cells and NIH-3T3 cells overexpressing insulin ( NIH-3T3(IR) ) or IGF-1 receptor ( NIH-3T3(IGF-1R) ) . Stimulation of insulin and IGF-1 receptors by insulin caused a temporary dissociation of annexin II from these receptors , which was accompanied by an increased amount of extracellular annexin II detected in the media of PC12 , NIH-3T3(IR) , and NIH-3T3(IGF-1R) cells but not in that of untransfected NIH-3T3 cells . Activation of a different growth factor receptor , the platelet-derived growth factor receptor , did not produce such results . Tyrphostin AG1024 , a tyrosine kinase inhibitor of insulin and IGF-1 receptor , was shown to inhibit annexin II secretion along with reduced receptor phosphorylation . Inhibitors of a few downstream signaling enzymes including phosphatidylinositol 3-kinase , pp60c-Src , and protein kinase C had no effect on insulin-induced annexin II secretion , suggesting a possible direct link between receptor activation and annexin II secretion . Immunocytochemistry revealed that insulin also induced transport of the membrane-bound form of annexin II to the outside layer of the cell membrane and appeared to promote cell aggregation . These results suggest that the insulin receptor and its signaling pathways may participate in molecular mechanisms mediating annexin II secretion . OUTPUT: sustaining proliferative signaling INPUT: Transforming growth factor beta ( TGF-β ) signaling has been implicated in driving tumor progression and metastasis by inducing stem cell-like features in some human cancer cell lines . In this study , we have utilized a novel murine cell line NMuMG-ST , which acquired cancer stem cell ( CSC ) phenotypes during spontaneous transformation of the untransformed murine mammary cell line NMuMG , to investigate the role of autocrine TGF-β signaling in regulating their survival , metastatic ability , and the maintenance of cancer stem cell characteristics . We have retrovirally transduced a dominant-negative TGF-β type II receptor ( DNRII ) into the NMuMG-ST cell to abrogate autocrine TGF-β signaling . The expression of DNRII reduced TGF-β sensitivity of the NMuMG-ST cells in various cell-based assays . The blockade of autocrine TGF-β signaling reduced the ability of the cell to grow anchorage-independently and to resist serum deprivation-induced apoptosis . These phenotypes were associated with reduced levels of active and phosphorylated AKT and ERK , and Gli1 expression suggesting that these pathways contribute to the growth and survival of this model system . More interestingly , the abrogation of autocrine TGF-β signaling also led to the attenuation of several features associated with mammary stem cells including epithelial-mesenchymal transition , mammosphere formation , and expression of stem cell markers . When xenografted in athymic nude mice , the DNRII cells were also found to undergo apoptosis and induced significantly lower lung metastasis burden than the control cells even though they formed similar size of xenograft tumors . Thus , our results indicate that autocrine TGF-β signaling is involved in the maintenance and survival of stem-like cell population resulting in the enhanced metastatic ability of the murine breast cancer cells . OUTPUT: resisting cell death;activating invasion and metastasis INPUT: Retinoic acid ( RA ) and its derivatives inhibit the proliferation of normal human mammary epithelial cells ( HMEC ) and some breast carcinoma lines by mechanisms which are not fully understood . To identify genes that mediate RA-induced cell growth arrest , an HMEC cDNA library was synthesized and subtractive screening was performed . We identified the interleukin-1beta ( IL-1beta ) gene as an RA induced gene in HMEC . Northern blot analyses showed that the IL-1beta gene was up-regulated as early as 2 h after RA treatment . Results from the treatment of HMEC with cycloheximide and actinomycin D indicated that the regulation of the IL-1beta gene by RA occurred at the transcriptional level and that the IL-1beta gene is a direct , downstream target gene of RA . To evaluate the effects of IL-1beta on cell proliferation , the proliferation of HMEC was measured in the presence of RA or IL-1beta , or both . Either RA or IL-1beta could significantly inhibit the proliferation of HMEC . However , the addition of soluble IL-1 receptor antagonist ( sIL-1ra ) to the cell culture medium did not block RA-induced HMEC growth inhibition , whereas sIL-1ra did block the growth inhibition of HMEC by IL-1beta . IL-1beta expression was not observed in the three carcinoma cell lines , MCF-7 , MDA-MB-231 , and MDA-MB-468 , as compared to the HMEC . Growth curves of the breast carcinoma cell lines showed strong inhibitory effects of RA and IL-1beta on the growth of the estrogen receptor ( ER ) positive MCF-7 cell line , but only a small effect on the ER negative MDA-MB-231 cells . The expression of the IL-1beta gene was also transcriptionally activated by RA in normal epithelial cells of prostate and oral cavity . Our results suggest that : ( a ) the IL-1beta gene is a primary target of RA receptors in HMEC ; ( b ) the enhanced expression of the IL-1beta gene does not mediate the RA-induced growth arrest of HMEC ; and ( c ) the expression of the IL-1beta gene is low or absent in all three human breast carcinoma cell lines examined , but the defect in the IL-1beta signaling pathway may be different in ER positive versus ER negative carcinoma cells . OUTPUT: sustaining proliferative signaling INPUT: Epidermal growth factor receptor-tyrosine kinase inhibitors ( EGFR-TKIs ) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor ( EGFR ) gene . On the other hand , some lung cancer patients with wild type EGFR also respond to EGFR-TKIs , suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells . However , the effect of EGFR-TKIs on host microenvironments is largely unknown . A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells . This model was used to investigate the therapeutic efficacy of erlotinib , an EGFR-TKI , on multiple organ metastases induced by human small cell lung cancer cells ( SBC-5 cells ) that did not express EGFR . Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro , it significantly suppressed bone and lung metastases in vivo , but not liver metastases . An immunohistochemical analysis revealed that , erlotinib significantly suppressed the number of osteoclasts in bone metastases , whereas no difference was seen in microvessel density . Moreover , erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line ( MC3T3-E1 cells ) . These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells . OUTPUT: activating invasion and metastasis;inducing angiogenesis;sustaining proliferative signaling INPUT: BACKGROUND Modulation of the expression of retinoic acid receptors ( RAR ) alpha and gamma in adult rat prostate by testosterone ( T ) suggests that RAR signaling events might mediate some of the androgen effects on prostate cells . METHOD In this study , we examined the interactions between T and retinoic acid ( RA ) in cell growth of human prostate carcinoma cells , LNCaP , and their relationship with the expression of RAR and epidermal growth factor receptor ( EGF-R ) . RESULTS Both T and RA , when administered alone , stimulated 3H-thymidine incorporation in LNCaP cells in a dose-dependent manner ; the effect of each agent was reciprocally attenuated by the other agent . Testosterone treatment of LNCaP cells also resulted in dose dependent , biphasic increases in RAR alpha and gamma mRNAs ; increases paralleled that of 3H-thymidine incorporation and were attenuated by the presence of 100 nM RA . These results suggest a link between RAR signaling and the effect of T on LNCaP cell growth . Gel electrophoretic mobility shift assays revealed the presence of putative androgen responsive element ( ARE ) in the promoter region of RAR alpha gene , suggesting that a direct AR-DNA interaction might mediate the effects of T on RAR alpha gene . Furthermore , treatment of LNCaP cells with 20 nM T resulted in an increase in EGF-R . In contrast , EGF-R was suppressed by 100 nM RA that also suppressed the effect of T. CONCLUSIONS Current results demonstrate interactions between T and RA in the expression of RARs and cell growth in LNCaP cells . The presence of putative ARE in the promoter of the RAR alpha gene suggests that AR-DNA interaction might mediate the effects of T on RAR alpha gene . The opposite effects of T and RA on the expression of RAR and EGF-R suggest that signal events of these receptors might be involved in the interaction between T and RA in the control of LNCaP cell growth . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot7

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: While human embryonic stem cells ( hESCs ) and human embryonal carcinoma cells ( hECCs ) have been studied extensively at the levels of the genome , transcriptome , proteome and epigenome our knowledge of their corresponding metabolomes is limited . Here , we present the metabolic signatures of hESCs and hESCs obtained by untargeted gas chromatography coupled to mass spectrometry ( GC-MS ) . Whilst some metabolites are common to both cell types , representing the self-renewal and house-keeping signatures , others were either higher ( e.g. , octadecenoic acid , glycerol-3-phosphate , 4-hydroxyproline ) or lower ( e.g. , glutamic acid , mannitol , malic acid , GABA ) in hESCs ( H9 ) compared to hECCs ( NTERA2 ) , these represent cell type specific signatures . Further , our combined results of GC-MS and microarray based gene expression profiling of undifferentiated and OCT4-depleted hESCs are consistent with the Warburg effect which is increased glycolysis in embryonic cells and tumor cells in the presence of O(2) while oxidative phosphorylation ( OXPHOS ) is impaired or even shut down . RNAi-based OCT4 knock down mediated differentiation resulted in the activation of the poised OXPHOS machinery by expressing missing key proteins such as NDUFC1 , UQCRB and COX , increase in TCA cycle activity and decreased lactate metabolism . These results shed light on the metabolite layer of pluripotent stem cells and could potentially establish novel metabolic markers of self renewal and pluripotency . OUTPUT: cellular energetics INPUT: Food-derived heterocyclic aromatic amines ( HCAs ) have proved to be carcinogenic in both rodents and nonhuman primates . Two different metabolic pathways are suggested for the metabolic activation of HCA . The hepatic pathway proceeds via a two-step process involving N-hydroxylation by cytochrome P4501A2 and subsequent O-acetylation by N-acetyltransferase-2 . An alternative pathway may be of particular interest in extrahepatic tissues and proceeds via one-electron oxidation catalyzed by prostaglandin H synthase ( PHS ) , rendering free-radical metabolites . In this study , we investigated the metabolic activation of two HCAs , 2-amino-3-methylimidazo[4,5-f]quinoline ( IQ ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , by two different enzyme systems in vitro , generating different primary and secondary reactive metabolites . Rat liver S9 mix and PHS were used as the activating system and represent the hepatic and extrahepatic pathways , respectively . Electron-spin resonance spectroscopy showed that both IQ and PhIP exerted inhibiting effects on PHS-mediated formation of hydroxyl radicals during the conversion of arachidonic acid to prostaglandins . Evidence for the formation of HCA free radicals was presented in an indirect way by the formation of glutathione-derived thiyl radicals , with purified PHS as the activating system . Activation by S9 mix did not result in the formation of detectable radical metabolites , showing that the two metabolic routes primarily led to the formation of different metabolites . In all electron-spin resonance experiments , IQ appeared to be more effective than PhIP . In contrasts , DNA adduct analysis by means of ( 32)P-postlabeling showed similar adduct patterns for S9 and PHS in single-stranded and double-stranded salmon testes DNA after incubation with PhIP , indicating the ultimate formation of a common reactive intermediate . For IQ , activation by PHS led to an additional adduct spot that was not present after S9 activation . Furthermore , activation of IQ resulted in higher adduct levels compared with PhIP for both activation pathways . Overall , adduct levels were higher in single-stranded DNA than double-stranded DNA . Our results showed that the hepatic and extrahepatic pathways resulted in different primary metabolites , while the ultimate formation of a similar reactive intermediate for PhIP , possibly an arylnitrenium ion , suggested that both pathways could play an important role in the onset of carcinogenesis . OUTPUT: genomic instability and mutation INPUT: The published results on nanoparticles cytotoxicity and genotoxicity such as titanium dioxide nanoparticles ( TiO(2) NPs ) are inconsistent , and often conflicting and insufficient . Since different parameters may have impact on the toxicity results , there is need to lay stress on detailed characterization of NPs and the use of different testing conditions for assessment of NPs toxicity . In order to investigate whether dispersion procedures influence NP cytotoxicity and genotoxicity , we compared two protocols giving TiO(2) NP dispersions with different stability and agglomeration states . Detailed primary and secondary characteristics of both TiO(2) NP dispersions in culture media were carried out before toxicological testing ; TK6 human lymphoblast cells , EUE human embryonic epithelial cells and Cos-1 monkey kidney fibroblasts were used to assess cytotoxicity ( by trypan blue exclusion , proliferation activity and plating efficiency assays ) and genotoxicity ( by the comet assay ) . DNA strand breaks were detected by the alkaline comet assay . DNA oxidation lesions ( especially 8-oxo-7,8-dihydroguanine , 8-oxoG ) were measured with a modified comet assay including incubation with specific repair enzyme formamidopyrimidine DNA glycosylase ( FPG ) . The TiO(2) NPs dispersion with large agglomerates ( 3 min sonication and no serum in stock solution ) induced DNA damage in all three cell lines , while the TiO(2) NPs dispersed with agglomerates less than 200 nm ( foetal serum in stock solution and sonication 15 min ) had no effect on genotoxicity . An increased level of DNA oxidation lesions detected in Cos-1 and TK6 cells indicates that the leading mechanism by which TiO(2) NPs trigger genotoxicity is most likely oxidative stress . Our results show that the dispersion method used can influence the results of toxicity studies . Therefore at least two different dispersion procedures should be incorporated into assessment of cyto- and genotoxic effects of NPs . It is important , when assessing the hazard associated with NPs , to establish standard testing procedures and thorough strategies to consider the diverse conditions relevant to possible exposures . OUTPUT: genomic instability and mutation;tumor promoting inflammation INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: The role of energy deregulation and altered/adapted metabolism in tumor cells is an increasingly important issue in understanding cancer . Hereditary leiomyomatosis and renal cell carcinoma ( HLRCC ) is an aggressive form of RCC characterized by germline mutation of fumarate hydratase ( FH ) , followed by somatic loss of the remaining wild-type allele and known to be a highly metastatic and lethal malignancy compared to other RCCs . The intrinsic loss of normal tricarboxylic acid ( TCA ) cycle presumably aids tumorigenesis due to the necessary metabolic alterations required and the enforced dependence on glycolysis derived energy , mimicking the Warburg effect . Thus , there is considerable utility in establishing a preclinical cell model from these tumors to study energy metabolism deregulation , as well as developing new targeted therapeutic approaches for TCA cycle enzyme-deficient cancers . Here , we describe a new immortalized cell line , UOK268 , derived from a patient's primary HLRCC-associated kidney cancer . This represents the first primary renal cell line to model TCA cycle gene loss and provides a perfect partner cell line to our previously described metastasis-derived HLRCC-associated cell line , UOK262 . We identified a novel germline FH missense mutation , p.His192Asp , and the subsequent loss of heterozygosity in UOK268 . The UOK268 cell line expressed mutant FH protein , which localized to the mitochondria , but with loss of almost all catalytic activity . The UOK268 cells had severely compromised oxidative phosphorylation and increased glycolytic flux . Ingenuity pathways analysis of human mitochondria-focused cDNA microarray ( hMitChip3 ) gene chip data confirmed the altered mRNA expression patterns of genes involved in several important pathways , such as lipid metabolism , apoptosis , and energy production/glycolysis . UOK268 provides a unique model of a primary cell line demonstrating an enforced , irreversible Warburg effect and , combined with UOK262 , provides a unique invitro preclinical model for studying the bioenergetics of the Warburg effect in human cancer . OUTPUT: genomic instability and mutation;cellular energetics;resisting cell death INPUT: The metabolic detoxification capacity may critically regulate the susceptibility of human tissues to cancer development . We used standardized and quantitative , reverse transcription-polymerase chain reaction ( StaRT-PCR ) and microarray chip techniques to analyze transcript levels of multiple detoxification enzymes in cultured normal human oral keratinocytes ( NOK ) and the Siman virus 40 T antigen-immortalized oral keratinocyte line SVpgC2a , viewing the latter as a model of a benign tumor state . With good agreement between the 2 methodologies , NOK and SVpgC2a were found to express transcripts for cytochrome P450 enzymes ( CYPs ) , factors related to CYP induction and enzymes involved in conjugation reactions or detoxification of reactive oxygen . The cell types expressed similar levels of CYP 2B6/7 , CYP 2E1 , P450 oxidoreductase , the aryl hydrocarbon receptor nuclear translocator , sulfotransferase 1A1 , sulfotransferase 1A3 , epoxide hydrolase , glutathione S-transferase M3 , glutathione S-transferase pi and catalase , superoxide dismutase 1 , glutathione peroxidase 1 and glutathione peroxidase 3 . In contrast , SVpgC2a exhibited comparatively higher levels of CYP1A1 , 1B1 , aryl hydrocarbon receptor , glutathione S-transferase M1 , 2 , 4 , 5 , glutathione S-transferase theta 1 and superoxide dismutase 2 and comparatively lower levels of UDP glycosyltransferase 2 and microsomal glutathione S-transferase 1 . Some transcripts , e.g. , CYP 2A6/7 , were not detected by either standard , non quantitative RT-PCR or the above methods , whereas others were barely quantifiable by StaRT-PCR , i.e. , were present at 1-10 molecules/10(6) molecules of actin . Overall , the expression analysis demonstrated presence of mRNA for multiple enzymes involved in foreign compound metabolism and detoxification pathways , including several enzymes not previously reported for oral epithelium . Generally , the comparison of NOK from 2 individuals indicated relatively similar transcript levels of these enzymes . In contrast , differences between NOK and SVpgC2a , e.g. , for CYP1B1 , may reflect alteration caused by immortalization and aid identification of early stage tumor markers in oral epithelium . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot8

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: OBJECTIVES To investigate the effects of serotonin and melatonin ( MLT ) on the regulation of malignant growth and the activity of serotonin receptors ( 5HTR1a/-1b ) in prostate cancer ( PCa ) cell lines . MATERIALS AND METHODS In four PCa cell lines ( LNCaP , 22RV1 , PC3 , DU145 ) and two reference cell lines 5HTR1a and -1b , relative mRNA expression levels were assessed . Different serotonin and MLT receptor agonists and antagonists were used in stimulation and inhibition experiments . RESULTS mRNA expression of 5HTR1b was higher than that of 5HTR1a in all PCa cell lines . Serotonin showed a significant growth stimulatory effect in all PCa lines . The 5HTR1a and -1b agonists/antagonists did not significantly affect viability . MLT inhibited viability only in PC3 cells . Similarly , the 5HTR1a antagonist induced apoptotic changes in PC3 cells only at 10(-4)M , while the 5HTR1b antagonist induced necrosis at 10(-4)M in all cell lines . Cell cycle alterations were seen in PC3 and DU145 cells under the influence of the 5HTR1a antagonist . CONCLUSIONS Serotonin receptor antagonists and agonists as well as MLT influence viability and apoptosis of PCa cell lines at supraphysiologic concentrations . In contrast to other reports , our results do not support a regulatory role of serotonin or MLT receptor activation or inhibition in PCa growth . OUTPUT: resisting cell death INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Energy deregulation and abnormalities of tumor cell metabolism are critical issues in understanding cancer . Hereditary leiomyomatosis renal cell carcinoma ( HLRCC ) is an aggressive form of RCC characterized by germline mutation of the Krebs cycle enzyme fumarate hydratase ( FH ) , and one known to be highly metastatic and unusually lethal . There is considerable utility in establishing preclinical cell and xenograft models for study of disorders of energy metabolism , as well as in development of new therapeutic approaches targeting of tricarboxylic acid ( TCA ) cycle enzyme-deficient human cancers . Here we describe a new immortalized cell line , UOK 262 , derived from a patient having aggressive HLRCC-associated recurring kidney cancer . We investigated gene expression , chromosome profiles , efflux bioenergetic analysis , mitochondrial ultrastructure , FH catabolic activity , invasiveness , and optimal glucose requirements for in vitro growth . UOK 262 cells have an isochromosome 1q recurring chromosome abnormality , i(1)(q10) , and exhibit compromised oxidative phosphorylation and in vitro dependence on anaerobic glycolysis consistent with the clinical manifestation of HLRCC . The cells also display glucose-dependent growth , an elevated rate of lactate efflux , and overexpression of the glucose transporter GLUT1 and of lactate dehydrogenase A ( LDHA ) . Mutant FH protein was present primarily in edematous mitochondria , but with catalytic activity nearly undetectable . UOK 262 xenografts retain the characteristics of HLRCC histopathology . Our findings indicate that the severe compromise of oxidative phosphorylation and rapid glycolytic flux in UOK 262 are an essential feature of this TCA cycle enzyme-deficient form of kidney cancer . This tumor model is the embodiment of the Warburg effect . UOK 262 provides a unique in vitro and in vivo preclinical model for studying the bioenergetics of the Warburg effect in human cancer . OUTPUT: cellular energetics INPUT: We investigated the direct effects of LH-releasing hormone ( LH-RH ) antagonist , Cetrorelix , on the growth of HTOA human epithelial ovarian cancer cell line . RT-PCR revealed the expression of mRNA for LH-RH and its receptor in HTOA cells . Cetrorelix , at concentrations between 10(-9) and 10(-5) M , exerted a dose-dependent antiproliferative action on HTOA cells , as measured by 5-bromo-2'-deoxyuridine incorporation into DNA . Flow cytometric analysis indicated that Cetrorelix , at 10(-5) M , arrested cell cycle in HTOA cells , at G1 phase , after 24 h of treatment . Western blot analysis of cell cycle-regulatory proteins demonstrated that treatment with Cetrorelix ( 10(-5) M ) for 24 h did not change the steady-state levels of cyclin D1 , cyclin E , and cyclin-dependent kinase ( Cdk)4 but decreased the levels of cyclin A and Cdk2 . The protein levels of p21 ( a Cdk inhibitor ) and p53 ( a suppressor of tumor cell growth and a positive regulator for p21 expression ) were increased by Cetrorelix , but the levels of p27 ( a Cdk inhibitor ) did not change significantly . Flow cytometric analysis and terminal deoxynucleotidyltransferase-mediated deoxyuridine 5-triphosphate nick end labeling staining demonstrated that Cetrorelix ( 10(-5) M ) induced apoptosis in HTOA cells . In conclusion , Cetrorelix directly inhibits the proliferation of human epithelial ovarian cancer cells through mechanisms mediated by LH-RH receptor and involving multiple events in cell cycle progression , including G1 phase cell cycle arrest coupled with down-regulation of cyclin A-Cdk2 complex levels , presumably attributable to an up-regulation of p53 and p21 protein levels and apoptosis . OUTPUT:

sustaining proliferative signaling;evading growth suppressors;resisting cell death

HoC_dynamic_5_shot9

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: The role of regulatory T cells ( T(regs) ) in human colon cancer ( CC ) remains controversial : high densities of tumor-infiltrating T(regs) can correlate with better or worse clinical outcomes depending on the study . In mouse models of cancer , T(regs) have been reported to suppress inflammation and protect the host , suppress T cells and protect the tumor , or even have direct cancer-promoting attributes . These different effects may result from the presence of different T(reg) subsets . We report the preferential expansion of a T(reg) subset in human CC with potent T cell-suppressive , but compromised anti-inflammatory , properties ; these cells are distinguished from T(regs) present in healthy donors by their coexpression of Foxp3 and RORγt . T(regs) with similar attributes were found to be expanded in mouse models of hereditary polyposis . Indeed , ablation of the RORγt gene in Foxp3(+) cells in polyp-prone mice stabilized T(reg) anti-inflammatory functions , suppressed inflammation , improved polyp-specific immune surveillance , and severely attenuated polyposis . Ablation of interleukin-6 ( IL-6 ) , IL-23 , IL-17 , or tumor necrosis factor-α in polyp-prone mice reduced polyp number but not to the same extent as loss of RORγt . Surprisingly , loss of IL-17A had a dual effect : IL-17A-deficient mice had fewer polyps but continued to have RORγt(+) T(regs) and developed invasive cancer . Thus , we conclude that RORγt has a central role in determining the balance between protective and pathogenic T(regs) in CC and that T(reg) subtype regulates inflammation , potency of immune surveillance , and severity of disease outcome . OUTPUT: tumor promoting inflammation INPUT: Cellular senescence forms a barrier that inhibits the acquisition of an immortal phenotype , a critical feature in tumorigenesis . The inactivation of multiple pathways that positively regulate senescence are required for immortalization . To identify these pathways in an unbiased manner , we performed DNA microarray analyses to assess the expression of 20,000 genes in human prostate epithelial cells ( HPECs ) passaged to senescence . These gene expression patterns were then compared with those of HPECs immortalized with the human Papillomavirus 16 E7 oncoprotein . Senescent cells display gene expression patterns that reflect their nonproliferative , differentiated phenotype and express secretory proteases and extracellular matrix components . A comparison of genes transcriptionally up-regulated in senescence to those in which expression is significantly down-regulated in immortalized HPECs identified three genes : the chemokine BRAK , DOC1 , and a member of the insulin-like growth factor axis , IGFBP-3 . Expression of these genes is found to be uniformly lost in human prostate cancer cell lines and xenografts , and previously , their inactivation was documented in tumor samples . Thus , these genes may function in novel pathways that regulate senescence and are inactivated during immortalization . These changes may be critical not only in allowing cells to bypass senescence in vitro but in the progression of prostate cancer in vivo . OUTPUT: enabling replicative immortality INPUT: To examine the association of cell cycle regulatory gene inactivation with human cell immortalization , we determined the expression status of INK4a , Rb , and WAF1/ CIP1 , in eleven in vitro immortalized human cell lines , including fibroblasts and keratinocytes . Two human papillomavirus type 16 E6 expressing cell lines with telomerase activity , including a fibroblast cell line and a keratinocyte cell line , expressed no detectable p16(INK4a) . These cell lines had a hyperphosphorylated pRb and reduced expression of p21(WAF1/CIP1) . All of seven fibroblast cell lines immortalized either spontaneously or by ( 60)Co , X-rays , 4-nitroquinoline 1-oxide or aflatoxin B(1) , maintaining their telomeres by the ALT ( alternative lengthening of telomeres ) pathway , displayed loss of expression of p16(INK4a) and hyperphosphorylation of pRb . Levels of p21(WAF1/CIP1) expression varied among the cell lines . Two fibroblast cell lines that became immortalized following infection with a retrovirus vector encoding human telomerase catalytic subunit ( hTERT ) cDNA were also accompanied by inactivation of p16(INK4a) and pRb pathways . Acquisition of telomerase activity alone was not sufficient for immortalization of these cell lines . Taken together , all the cell lines including fibroblasts and keratinocytes , with either telomerase activity or the ALT pathway for telomere maintenance showed loss of expression of p16(INK4a) and hyperphosphorylation of pRb . These demonstrate the association of inactivation of both p16(INK4a) and pRb with immortalization of human cells including fibroblasts and epithelial cells and telomerase-positive cells and ALT-positive cells . OUTPUT: enabling replicative immortality INPUT: Direct experimental evidence implicates telomere erosion as a primary cause of cellular senescence . Using a well characterized model system for breast cancer , we define here the molecular and cellular consequences of adriamycin treatment in breast tumor cells . Cells acutely exposed to adriamycin exhibited an increase in p53 activity , a decline in telomerase activity , and a dramatic increase in beta-galactosidase , a marker of senescence . Inactivation of wild-type p53 resulted in a transition of the cellular response to adriamycin treatment from replicative senescence to delayed apoptosis , demonstrating that p53 plays an integral role in the fate of breast tumor cells treated with DNA-damaging agents . Stable introduction of hTERT , the catalytic protein component of telomerase , into MCF-7 cells caused an increase in telomerase activity and telomere length . Treatment of MCF-7-hTERT cells with adriamycin produced an identical senescence response as controls without signs of telomere shortening , indicating that the senescence after treatment is telomere length-independent . However , we found that exposure to adriamycin resulted in an overrepresentation of cytogenetic changes involving telomeres , showing an altered telomere state induced by adriamycin is probably a causal factor leading to the senescence phenotype . To our knowledge , these data are the first to demonstrate that the mechanism of adriamycin-induced senescence is dependent on both functional p53 and telomere dysfunction rather than overall shortening . OUTPUT: enabling replicative immortality;genomic instability and mutation;resisting cell death INPUT: Id proteins are negative regulators of basic helix-loop-helix transcription factors , which are critical for expression of genes associated with cellular differentiation . Previous studies have shown that overexpression of Id-1 delays cellular senescence in several cell types , including fibroblasts , mammary epithelial cells , and keratinocytes . Although previous studies have demonstrated the expression of Id-1 in endothelium , the regulation of Id-1 has not been studied in these cells . In this report , a retroviral vector was used to overexpress Id-1 in human endothelial cells . Sustained expression of Id-1 resulted in a 2- to 3-fold increase in the total number of population doublings ( replicative capacity ) of the cells compared with vector-treated controls , which correlated with low levels of p16 , p21 , and p27 expression . The cells , however , were not immortalized and did eventually undergo senescence despite elevated Id-1 levels . Senescence was characterized by a dramatic increase in p16 , but not p21 and p27 . Under these experimental conditions , telomerase activity was not detected and the telomeres became progressively shorter with time . These results demonstrate the importance of Id-1 in endothelial cell proliferation and indicate that Id-1 represses p16 expression , resulting in delayed senescence . These findings may have implications in the development of endothelial cell-derived tumors . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot10

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: The role of T cells in eradicating leukemic cells has been well demonstrated for chronic myeloid leukemia ( CML ) . Type 1 ( T1 ) T-cell cytokines play a major role in this antileukemic immune effect . Studies in cancer patients have demonstrated a decreased T1 cytokine production , measured by enzyme-linked immunosorbent assay ( ELISA ) , in cultures of peripheral blood mononuclear cells . This observation of malignancy-related suppressed T1 cytokines also occurs in untreated chronic-phase ( CP ) CML , raising the question of the influence of different CML treatment regimens on this immunosuppression . Intracellular flow cytometry ( ICF ) has facilitated the evaluation of cytokines on a single-cell level . This study analyzed T1 ( interferon-gamma ) cytokine production in purified peripheral blood T cells by ICF , comparing different therapy approaches for CML . Twenty-one newly diagnosed CP CML patients were compared with 24 patients treated with interferon-alpha ( IFN-alpha ) and to 30 allogeneic bone marrow transplant ( BMT ) recipients ( BCR-ABL negative by reverse-transcriptase polymerase chain reaction , and free of , or having only limited graft-versus-host disease at the time of study ) . Thirty-seven healthy controls were included . Our results showed a significantly decreased T-cell IFN-gamma synthesis in CP CML patients in relation to healthy controls ( P = 0.0007 ) . Treatment with IFN-alpha resulted in a shift from immunosuppression--documented for the group of untreated patients--to immunopotentiation , with an increase of T-cell IFN-gamma production ( P = 0.0266 ) . Notably , BMT enhanced IFN-gamma production of T cells to a level not only exceeding untreated patients ( P &lt ; 0.0001 ) but also healthy volunteers ( P &lt ; 0.0001 ) . The observation of T1 cytokine up-regulation with IFN-alpha therapy indicates that enhanced T-cell function may be achievable in patients with CML , even in the absence of an allo-response . OUTPUT: avoiding immune destruction INPUT: Interferon-gamma ( IFN-gamma ) has pleiotropic activities other than its antivirus action , including cell growth inhibition , natural killer ( NK ) cell and cytotoxic T lymphocyte ( CTL ) activation , and angiogenesis inhibitory activity , and these activities are supposed to be involved in its antitumour activity . However , it has not been completely elucidated which activity is mainly involved in the tumour suppression in vivo . In this study , we analysed inhibitory mechanisms of endogenous IFN-gamma against B16 melanoma experimental metastasis . After intravenous injection of tumour cells , tumour deposits in the lungs and liver were increased and life span was shorter in IFN-gamma(-/-) mice , indicating important roles for IFN-gamma in antitumour mechanisms . Interestingly , tumour deposits were not increased in IFN-gamma receptor ( R)(-/- ) mice . Furthermore , only low levels of cell-mediated immunity against the tumour and activation of NK cells were observed , indicating that antimetastatic effects of IFN-gamma is not mediated by host cells . The survival period of B16 melanoma-bearing IFN-gamma R(-/-) mice was , however , shorter than wild-type mice . These observations suggest that IFN-gamma prevents B16 melanoma experimental metastasis by directly inhibiting the cell growth , although antitumour host functions may also be involved in a later phase . OUTPUT: activating invasion and metastasis INPUT: It has been shown that injecting a suspension of IFN-γ-secreting tumor cells results in their rejection . This effect has been attributed to IFN-γ preventing tumor stroma formation but not to a direct effect on the cancer cells . However , it is not known , which influence IFN-γ has on tumors with an established stroma . To address this question , the plasmacytoma cell line J558L was transduced with a vector allowing doxycycline-inducible IFN-γ gene expression . After the injection of the tumor cells into mice , IFN-γ was induced at different time points . Tumors did not grow when inducing IFN-γ immediately after tumor cell inoculation , while approximately half of the tumors were rejected when IFN-γ was induced in early established tumors within 2 weeks . Induction of IFN-γ 2-3 weeks after tumor cell inoculation was less efficient ( 0-17% rejection ) . IFN-γ induction in established tumors led to a reduction of CD146(+) endothelial cells and massive necrosis . Together , we show that vascularized tumors can be rejected by local IFN-γ expression , but that rejection of established tumors was less efficient over time . This suggests that transplanted tumors became less susceptible to local IFN-γ treatment the better they are established . OUTPUT: resisting cell death;inducing angiogenesis INPUT: An efficent antitumor and antiviral cellular immune response requires optimal interferon-gamma ( IFN-gamma ) secretion and perforin expression in CD8(+) T cells . The aim of this study was to define whether CD4(+) and CD8(+) T cells from patients with undifferentiated carcinoma of nasopharyngeal type ( UCNT ) , a tumor regularly associated with the Epstein-Barr virus ( EBV ) , have abnormal phenotype profiles , cytokine production , perforin and CD3-zeta expressions . Our data showed that CD4 and CD8 subset distribution was not grossly altered in the peripheral blood of UCNT patients , while tumor biopsies contained an increased proportion of CD8(+) T cells . The analysis of the CD4(+) subset showed a defect in interleukin-2 ( IL-2 ) production and a moderate increase of IL-10 production , a situation consistent with a Th1/Th2 imbalance . We have also demonstrated that CD8(+) lymphocytes from UCNT patients had a marked impairment of IFN-gamma secretion and perforin expression . This impairment was not related to the presence of detectable EBV DNA in the plasma . In UCNT patients , the blockade of the perforin pathway and of IFN-gamma production may constitute important mechanisms for immune escape by the tumor and for impaired control of EBV replication . OUTPUT: avoiding immune destruction INPUT: Blind mole rats Spalax ( BMR ) are small subterranean rodents common in the Middle East . BMR is distinguished by its adaptations to life underground , remarkable longevity ( with a maximum documented lifespan of 21 y ) , and resistance to cancer . Spontaneous tumors have never been observed in spalacids . To understand the mechanisms responsible for this resistance , we examined the growth of BMR fibroblasts in vitro of the species Spalax judaei and Spalax golani . BMR cells proliferated actively for 7-20 population doublings , after which the cells began secreting IFN-β , and the cultures underwent massive necrotic cell death within 3 d . The necrotic cell death phenomenon was independent of culture conditions or telomere shortening . Interestingly , this cell behavior was distinct from that observed in another long-lived and cancer-resistant African mole rat , Heterocephalus glaber , the naked mole rat in which cells display hypersensitivity to contact inhibition . Sequestration of p53 and Rb proteins using SV40 large T antigen completely rescued necrotic cell death . Our results suggest that cancer resistance of BMR is conferred by massive necrotic response to overproliferation mediated by p53 and Rb pathways , and triggered by the release of IFN-β . Thus , we have identified a unique mechanism that contributes to cancer resistance of this subterranean mammal extremely adapted to life underground . OUTPUT: resisting cell death;enabling replicative immortality;evading growth suppressors INPUT: When cells were incubated in the presence of both interferon-gamma ( IFN-gamma ) and all-trans retinoic acid ( ATRA ) , the concentration of IFN-gamma required to induce apoptosis of B-cell lymphoma cells was much lower than that required for myeloid or erythroid cell lines . The concentration of IFN-gamma that effectively inhibited the proliferation of BALM-3 cells was 1/40 of that required for BALM-1 cells . STAT-1 phosphorylation , IRF-1 mRNA and protein expression and RAR-beta expression were enhanced to a greater degree in BALM-3 cells treated with IFN-gamma and ATRA than in BALM-1 cells treated with IFN-gamma and ATRA , suggesting that these IFN-gamma related genes were involved in the induction of apoptosis of BALM-3 cells . OUTPUT:

resisting cell death

HoC_dynamic_5_shot11

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cell invasion is required for neoplastic metastasis . Matrix metalloproteinase-9 ( MMP-9 ) , which degrades the extracellular matrix , is a major component in the process of cancer cell invasion . Sulfuretin is one of the major flavonoids isolated from Rhus verniciflua . Sulfuretin has been used to reduce oxidative stress , platelet aggregation , the inflammatory response and mutagenesis . However , the effect of sulfuretin on breast cancer metastasis is unknown . In this study , we investigated the inhibitory effect of sulfuretin on 12-O-tetradecanoylphorbol-13-acetate ( TPA)-induced MMP-9 expression and cell invasion in MCF-7 cells . Sulfuretin inhibited TPA-induced transcriptional activation of nuclear factor-κB ( NF-κB ) . We demonstrated that sulfuretin mediated the inhibition of TPA-induced MMP-9 expression and that cell invasion in MCF-7 cells involved suppression of the NF-κB pathway . Therefore , inhibiting MMP-9 expression by sulfuretin may have therapeutic potential for controlling breast cancer invasiveness . OUTPUT: activating invasion and metastasis INPUT: Metastasis is a major cause of death of patients with malignant tumors . Matrix metalloproteinases ( MMPs ) are important for the migration and invasion of various types of cancer cell . Propofol is a known anesthetic agent , widely used for short-term anesthesia and for longer-term sedation . Propofol inhibits the proliferation of a variety of tumor cells , but there is no available information regarding propofol-inhibited migration and invasion of tumor cells in vitro . In this study , we investigated the effects of propofol on the migration and invasion of human lung carcinoma A549 cells . Wound healing assay and Boyden chamber assays indicated that propofol inhibited the migration and invasion of A549 cells in vitro . Gelatin zymographic analysis showed the inhibitory effect of propofol on the activation of expression MMP-2 . Western blot analysis also indicated that propofol suppressed the protein expiration of growth factor receptor-bound protein 2 ( GRB2 ) , Jun N-terminal kinases 1/2 ( p-JNK1/2 ) , p-p38 , MMP-2 and MMP-9 in A549 cells . Results from real-time PCR assay also showed that propofol inhibited the mRNA gene expression of MMP-2 , -7 and -9 , and enhanced that of tissue inhibitor of metalloproteinase 1 ( TIMP1 ) and TIMP2 in A549 cells . Taken together , these data show that propofol inhibits MMP-2 and -9 mRNA and protein expressions , resulting in suppression of lung cancer cell invasion and migration in vitro . OUTPUT: activating invasion and metastasis INPUT: High-grade prostate cancers express high levels of matrix metalloproteinases ( MMPs ) , major enzymes involved in tumor invasion and metastasis . However , the tumor cell lines commonly employed for prostate cancer research express only small amounts of MMPs when cultivated as monolayer cultures , in common culture media . The present study was conducted to ascertain whether culture conditions that include fibronectin can alter MMP2 and MMP9 expression by the human prostatic epithelial cell lines RWPE-1 , LNCaP and PC-3 . These cells were individually seeded at 2×10(4) cells/cm(2) , cultivated until they reached 80% confluence , and then exposed for 4h to fibronectin , after which the conditioned medium was analyzed by gelatin zymography . Untreated cells were given common medium . Only RWPE-1 cells express detectable amounts of MMP9 when cultivated in common medium , whereas the addition of fibronectin induced high expression levels of pro and active forms of MMP2 in all tested cell lines . Our findings demonstrate that normal and tumor prostate cell lines express MMP2 activity when in contact with extracellular matrix components or blood plasma proteins such as fibronectin . Future studies of transcriptomes and proteomes in prostate cancer research using these cell lines should not neglect these important conclusions . OUTPUT: activating invasion and metastasis INPUT: OBJECTIVE To observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe ( RSR ) on HER2 positive breast cancer , to evaluate the effects of RSR on the expressions of matrix metalloproteinases ( MMPs ) and the tissue inhibitor of metalloproteinases ( TIMPs ) in the recurrence and metastasis of HER2 positive breast cancer , thus revealing its anti-tumor recurrent and metastatic mechanisms . METHODS Selected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu . The primary tumor resection was carried out . After surgery they were randomly divided into the blank control group , the RSR group , the Herceptin group , and the combination group ( RSR + Herceptin group ) . The treatment lasted for 4 months . The inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated . The expressions of matrix metalloproteinase-2 ( MMP-2 ) , matrix metalloproteinase-9 ( MMP-9 ) , tissue inhibitor of metalloproteinase ( TIMP-1 ) , and TIMP-2 in the recurrent tumor tissue were detected using Western blot . RESULTS By the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group , showing statistical difference between the two groups ( P = 0.037 ) . The average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group . The inhibition rate of lung metastasis was 37. 85% in the RSR group , but with no statistical significance . The expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group , the Herceptin group , and the combination group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group , the Herceptin group , and the combination group was significantly down-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group ( P &lt ; 0.05 ) . The expressions of both TIMP-1 and TIMP-2 of the RSR group , the Herceptin group , and the combination group were all up-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group ( P &lt ; 0.05 ) . It was higher in the combination group than in the RSR group ( P &lt ; 0.05 ) . CONCLUSIONS RSR could inhibit the tumor recurrence of FVB/neu mice . It could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix . It might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND/AIMS We investigated whether the anticancer drug Ukrain ( UK ) is able to modulate the expression of some of the key markers of tumor progression in pancreatic cell carcinoma , in order to assess its potential therapeutic effect . METHODS Three cell lines ( HPAF-II , PL45 , HPAC ) were treated with UK ( 5 , 10 and 20 μM ) for 48 h , or left untreated . Secreted protein acidic and rich in cysteine ( SPARC ) mRNA levels were assessed by real-time PCR . Matrix metalloproteinases ( MMP)-2 and -9 activity was analyzed by SDS zymography ; SPARC protein levels in cell lysates and supernatants were determined by Western blot . Cell cycle was determined by flow cytometric analysis , and invasion by matrigel invasion assay . RESULTS UK down-regulated MMP-2 and MMP-9 , suggesting that UK may decrease pancreatic cancer cell invasion , as confirmed by the matrigel invasion assay . SPARC protein down-regulation in supernatants points to an inhibition by UK of extracellular matrix remodeling in the tumor microenvironment . At the same time , SPARC mRNA and cellular protein level up-regulation suggests that UK can affect cell proliferation by cell cycle inhibition , showing a cell cycle G2/M arrest in UK-treated cells . CONCLUSION Our results suggest that UK modulates two major aspects involved in tumorigenesis of pancreatic cancer cells , such as extracellular matrix remodeling and cell proliferation . OUTPUT: activating invasion and metastasis;evading growth suppressors INPUT: BACKGROUND Thrombospondin-1 ( TSP-1 ) promotes breast cancer cell invasion of collagen by upregulating matrix metalloproteinase-9 ( MMP-9 ) production . Stromal TSP-1 may play a role in regulating tumor cell invasion . We hypothesize that fibroblasts promote breast cancer cell invasion by upregulating the production of MMP-9 through TSP-1 . METHODS MDA-MB-231 human breast carcinoma cells were grown alone or in coculture with human fibroblasts . Gelatin zymography and Western immunoblot analysis for MMP-9 were performed on the coculture cell media and the single cell media . Inhibition of fibroblast-mediated breast tumor cell invasion by an anti-TSP-1 or an anti-MMP-9 antibody was evaluated using a modified Boyden chamber . RESULTS Coculture experiments showed an increased production of MMP-9 when compared with breast cancer single cell culture or fibroblast single cell culture experiments as demonstrated by zymography and Western immunoblot analysis . Fibroblast-stimulated MMP-9 production was comparable with TSP-1-stimulated MMP-9 production . Anti-TSP-1 antibody and anti-MMP-9 antibody inhibited fibroblast-stimulated tumor cell invasion to 30% and 26% of controls , respectively ( P <.05 ) . CONCLUSIONS Fibroblasts may regulate breast cancer cell invasion by promoting tumor MMP-9 production through TSP-1 . Inhibition of stromal TSP-1 stimulation of MMP-9 synthesis may prevent matrix degradation necessary for tumor invasion and metastasis . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot12

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: A new cell line of human ovarian clear cell adenocarcinoma ( CCC ) , TU-OC-1 , was established and characterized . The cells showed a polygonal-shaped morphology and grew in monolayers without contact inhibition and were arranged like a jigsaw puzzle . The chromosome numbers ranged from 64 to 90 . A low rate of proliferation was observed , similar to other CCC cell lines tested ( OVTOKO , RMG-I , and OVAS ) , and the doubling time was 38.4h . The respective IC50 values of cisplatin and paclitaxel were 12.2μM and 58.3nM . Mutational analysis revealed that TU-OC-1 cells harbored a PIK3CA mutation at codon 542 ( E542K ) in exon 9 , which is a mutation hot spot on this gene . We observed that phosphorylated Akt protein was overexpressed in TU-OC-1 cells by western blot analysis . Heterotransplantation to nude mice produced tumors that reflected the original . This cell line may be useful to study the chemoresistant mechanisms of CCC and contribute to novel treatment strategies . OUTPUT: genomic instability and mutation INPUT: UNLABELLED BACKGROUND Cell lines are very useful for clinical and basic research . Thus far , only 11 reports have documented the characteristics of ovarian endometrioid adenocarcinoma cell lines in the literature . Due to the scarcity of information , the establishment of an ovarian malignant tumor cell line with distinctive characteristics is particularly important to study this disease . Thus , this study was undertaken to establish and characterize a new human endometrioid adenocarcinoma cell line of the ovary . METHODS The cell line NOMH-1 was established from an ovarian tumor of a 44-year-old woman . Features of the cell line studied included morphology , chromosome analysis , heterotransplantation , tumor markers , and chemosensitivity . RESULTS This cell line has been growing well for 232 months and subcultured more than 50 times . Monolayer cultured cells were polygonal in shape , showing a pavement-like arrangement and a tendency to stack without contact inhibition . They exhibited a human karyotype with a modal chromosomal number in the hypertriploid range . The cells could be transplanted into the subcutis of nude mice and produced tumors resembling the original tumor . NOMH-1 cells expressed both CEA and CA19-9 which were identified immunohistochemically in the original tumor and the heterotransplanted tumor . The cells were sensitive to paclitaxel , an agent commonly used in the treatment of gynecological cancers . CONCLUSIONS NOMH-1 is the first ovarian endometrioid adenocarcinoma cell line in which CEA and CA19-9 expression have been defined . This newly established cell line should be useful for investigating the characteristics of ovarian endometrioid adenocarcinoma . OUTPUT: evading growth suppressors INPUT: We present a new cell line , EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient . The cells show rapid growth in culture with a doubling time of 16 h and high migration activity . Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition . Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma , whereas no metastasis was observed . Cultured EJ cells produced tissue polypeptide antigen ( IPA ) . Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression . Using the DNA sequencing technique , we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed . This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior , and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium . OUTPUT: activating invasion and metastasis;evading growth suppressors;sustaining proliferative signaling;genomic instability and mutation INPUT: BACKGROUND Human Barrett's cancer cell lines have numerous , poorly-characterized genetic abnormalities and , consequently , those lines have limited utility as models for studying the early molecular events in carcinogenesis . Cell lines with well-defined genetic lesions that recapitulate various stages of neoplastic progression in Barrett's esophagus would be most useful for such studies . METHODOLOGY/PRINCIPAL FINDINGS To develop such model cell lines , we started with telomerase-immortalized , non-neoplastic Barrett's epithelial ( BAR-T ) cells , which are spontaneously deficient in p16 , and proceeded to knock down p53 using RNAi , to activate Ras by introducing oncogenic H-Ras(G12V) , or both . BAR-T cells infected with either p53 RNAi or oncogenic H-Ras(G12V) alone maintained cell-to-cell contact inhibition and did not exhibit anchorage-independent growth in soft agar . In contrast , the combination of p53 RNAi knockdown with expression of oncogenic H-Ras(G12V) transformed the p16-deficient BAR-T cells , as evidenced by their loss of contact inhibition , by their formation of colonies in soft agar , and by their generation of tumors in immunodeficient mice . CONCLUSIONS/SIGNIFICANCE Through these experiments , we have generated a number of transformed and non-transformed cell lines with well-characterized genetic abnormalities recapitulating various stages of carcinogenesis in Barrett's esophagus . These lines should be useful models for the study of carcinogenesis in Barrett's esophagus , and for testing the efficacy of chemopreventive and chemotherapeutic agents . OUTPUT: evading growth suppressors INPUT: A new cell line , CB109 , has been established from a human glioblastoma multiforme . The cytoskeleton was positive for glial fibrillary acidic protein , vimentin and fibronectin . Hyaluronan ( HA ) and the HA-binding protein hyaluronectin ( HN ) were expressed in the cell cytoplasm and in the extracellular matrix of spheroids and plated cells . Hyaluronidase did not prevent spheroid formation suggesting that HA was not involved in the cell-cell adhesion . HA precoating prevented cell adherence to the plates and favoured spheroid formation . HA was secreted in relatively large amounts into the culture medium . High performance liquid chromatography demonstrated that HA was in the high molecular weight form . The rate of HN secretion by cells was very low . Basic fibroblast growth factor significantly increased the proliferation in vitro and tumour growth after grafting into nude mice . The epidermal growth factor receptor was not expressed on cultivated CB109 cells . Cytogenetic analysis showed polysomy 7 , structural rearrangement of chromosome 10 short arm and a translocation 13q13-q14 without detectable alteration of the RB gene . OUTPUT: sustaining proliferative signaling INPUT: A cell line designated HUUCLEC was established from a human uterine cervical lymphoepithelial carcinoma obtained from a 61-year-old Japanese woman . The cell line has grown slowly without interruption and serial passages were successively carried out 60 times within 3 years . The cultured cells were spindle or round in shape , showing anaplastic and pleomorphic features , a pavement cell arrangement and multilayering without contact inhibition . The population doubling time of the HUUCLEC line was 72 hours while the chromosomal number varied widely and showed aneuploidy . The modal chromosomal number was stable at the triploid range and marker chromosomes were present ; the Ebstein-Barr virus was absent in the cultured cells . OUTPUT:

evading growth suppressors;genomic instability and mutation

HoC_dynamic_5_shot13

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers . A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation . We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products . This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine ( NMBzA ) , and the response that increased levels of dietary vitamin E has on this induced carcinogenesis . Female C57BL/6 mice received NMBzA or vehicle ( corn oil ) i.p. weekly for 3 weeks . Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet . As an assessment of free radical activity , exhaled ethane was measured prior to killing the animals at 26 weeks . Esophagi from the various mice groups were assessed for size and frequency of tumors . Livers homogenates were analyzed for vitamins A and E , lipid fluorescence , conjugated dienes and malondialdehyde . Hepatic levels of vitamin A and E were decreased ( P &lt ; 0.05 ) and indices of lipid peroxidation were greater ( P &lt ; 0.05 ) in NMBzA-treated mice relative to controls . Lipid peroxidation and serum transaminases ( ALT and AST ) were greatest in mice given NMBzA and infected with the retroviruses . Incidence of esophageal tumors were also greatest in the NMBzA-treated , immunocompromised animals . Mice fed vitamin E-supplemented diets showed increased ( P &lt ; 0.05 ) hepatic concentrations of vitamin E and vitamin A , decreased activities of serum transaminases , decreased indices of lipid peroxidation , and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice . These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals . OUTPUT: avoiding immune destruction INPUT: Immunosuppression has been related to the incidence of tumor apparition , including endocrine tumors . The intrasplenic ovarian tumor ( luteoma ) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and , from the immunological perspective , is located in a critical organ involved in immune response . To establish if immunosuppression could alter the development of this experimental tumor , the effects of cyclosporin A ( CsA ) and dexamethasone ( Dex ) were evaluated . After surgery , tumor-bearing and sham animals were kept without treatment for 4 weeks ; thereafter , they were distributed into CsA ( 25 mg/kg ) , Dex ( 0.1 mg/kg ) , or vehicle ( 75:25 castor oil:ethanol ) groups and were injected on alternate days for 50 days . Body weight was evaluated weekly . Animals were sacrificed after a jugular vein blood sample was obtained . Thymi were weighed . Tumors were measured and placed in formaline for histological studies . Serum luteinizing hormone ( LH ) , follicle-stimulating hormone ( FSH ) , prolactin ( PRL ) , and estradiol were measured by radioimmunoassay . Hematological parameters were determined . CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals ( P &lt ; 0.01 ) . Dex significantly impaired weight increase in both groups of animals . CsA induced a significant weight loss in sham animals , not observed in tumor-bearing animals . Dex induced thymus weight loss in both groups , whereas CsA induced thymus weight loss only in sham animals . Only Dex induced a decrease in lymphocyte number in both groups . CsA induced an increase in monocyte number only in sham animals . Treatments did not alter LH , FSH , or estradiol , whereas PRL was increased by CsA only in sham rats . Neither Dex nor CsA induced any significant variations in tumor volume , nor did they alter tumor histology . In addition , no visible metastases or alterations in other organs were observed . We conclude that , though immunological parameters were altered by the treatments , immunosuppressor drugs did not condition tumor development . In addition , tumors secrete one or more factor/s that counteract CsA effect . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: The SR/CR mouse phenotype , first described in 1999 in BALB/c and later bred into C57BL/6 mice , is resistant to cancer formation following high doses of cancer cells administered intraperitoneally . The tumor cell targeting and destruction mechanisms have not been identified . By fluorescence-activated cell sorting analysis , the immune response of SR/CR mice after intraperitoneal injection of cancer cells was investigated and compared with parent strain mice . A massive influx of leukocytes into the peritoneal cavity was found . A large fraction of these leukocytes were polymorphonuclear granulocytes , macrophages and natural killer cells . A relative decrease in influx of B-cells compared with controls was demonstrated . Increased proportions of leukocytes belonging to the innate immune system were also demonstrated in splenocytes of SR/CR mice . Cytospins of peritoneal fluid from SR/CR mice after cancer cell injection showed formations of immune cells morphologically resembling polymorphonuclear granulocytes and macrophages adjoining the cancer cells . The results point to the potential involvement of innate immune cells in cancer immunology . Our data support migration of polymorphonuclear granulocytes , macrophages and NK cells into the peritoneum of the SR/CR mouse in response to intraperitoneal injection of S180 cancer cells . The cell composition of spleens of SR/CR mice reflected the differential regulation of the innate immune cells in peritoneal exudates . Both peritoneal exudates and the spleens of SR/CR mice contained decreased proportions of B-cells compared with BALB/c and C57BL/6 mice . We reproduce important aspects of previous published data and further extend them by showing differentially regulated populations of splenocytes including B-lymphocytes in SR/CR mice compared with parent strain controls . Importantly , this differentially regulated immune response of SR/CR mice could not be found in response to challenge with the lymphoma cell line EL-4 . OUTPUT: avoiding immune destruction INPUT: BACKGROUND Death receptors ( DR ) of the TNF family function as anti-tumor immune effector molecules . Tumor cells , however , often exhibit DR-signaling resistance . Previous studies indicate that radiation can modify gene expression within tumor cells and increase tumor cell sensitivity to immune attack . The aim of this study is to investigate the synergistic effect of sub-lethal doses of ionizing radiation in sensitizing colorectal carcinoma cells to death receptor-mediated apoptosis . METHODOLOGY/PRINCIPAL FINDINGS The ability of radiation to modulate the expression of multiple death receptors ( Fas/CD95 , TRAILR1/DR4 , TRAILR2/DR5 , TNF-R1 and LTβR ) was examined in colorectal tumor cells . The functional significance of sub-lethal doses of radiation in enhancing tumor cell susceptibility to DR-induced apoptosis was determined by in vitro functional sensitivity assays . The longevity of these changes and the underlying molecular mechanism of irradiation in sensitizing diverse colorectal carcinoma cells to death receptor-mediated apoptosis were also examined . We found that radiation increased surface expression of Fas , DR4 and DR5 but not LTβR or TNF-R1 in these cells . Increased expression of DRs was observed 2 days post-irradiation and remained elevated 7-days post irradiation . Sub-lethal tumor cell irradiation alone exhibited minimal cell death , but effectively sensitized three of three colorectal carcinoma cells to both TRAIL and Fas-induced apoptosis , but not LTβR-induced death . Furthermore , radiation-enhanced Fas and TRAIL-induced cell death lasted as long as 5-days post-irradiation . Specific analysis of intracellular sensitizers to apoptosis indicated that while radiation did reduce Bcl-X(L) and c-FLIP protein expression , this reduction did not correlate with the radiation-enhanced sensitivity to Fas and/or TRAIL mediated apoptosis among the three cell types . CONCLUSIONS/SIGNIFICANCE Irradiation of tumor cells can overcome Fas and TRAIL resistance that is long lasting . Overall , results of these investigations suggest that non-lethal doses of radiation can be used to make human tumors more amenable to attack by anti-tumor effector molecules and cells . OUTPUT: resisting cell death INPUT: The ultraviolet radiation present in sunlight is immune suppressive . Recently we showed that solar-simulated ultraviolet radiation ( ultraviolet A + B ; 295-400 nm ) , applied after immunization , suppressed immunologic memory and the elicitation of delayed-type hypersensitivity to the common opportunistic pathogen , Candida albicans . Further , we found that wavelengths in the ultraviolet A region of the solar spectrum ( 320-400 nm ) , devoid of ultraviolet B , were equally effective in activating immune suppression as ultraviolet A + B radiation . Here we report on the mechanisms involved . Maximal immune suppression was found when mice were exposed to solar-simulated ultraviolet radiation 7-9 d post immunization . No immune suppression was found in ultraviolet-irradiated mice injected with monoclonal anti-interleukin-10 antibody , or mice exposed to solar-simulated ultraviolet radiation and injected with recombinant interleukin-12 . Suppressor lymphocytes were found in the spleens of mice exposed to ultraviolet A + B radiation . In addition , antigen-specific suppressor T cells ( CD3+ , CD4+ , DX5+ ) were found in the spleens of mice exposed to ultraviolet A radiation . Applying liposomes containing bacteriophage T4N5 to the skin of mice exposed to solar-simulated ultraviolet A + B radiation , or mice exposed to ultraviolet A radiation , blocked immune suppression , demonstrating an essential role for ultraviolet-induced DNA damage in the suppression of established immune reactions . These findings indicate that overlapping immune suppressive mechanisms are activated by ultraviolet A and ultraviolet A + B radiation . Moreover , our findings demonstrate that ultraviolet radiation activates similar immunologic pathways to suppress the induction of , or the elicitation of , the immune response . OUTPUT:

avoiding immune destruction;genomic instability and mutation

HoC_dynamic_5_shot14

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Alterations of the epidermal growth factor receptor ( EGFR ) gene are common in some forms of cancer and the most frequent is a deletion of exons 2-7 . We have previously shown that this mutant receptor , called DeltaEGFR , confers enhanced tumorigenicity to glioblastoma cells through elevated proliferation and reduced apoptotic rates of the tumor cells in vivo . To understand the molecular mechanisms that underlie DeltaEGFR-enhanced proliferation , we examined the gene products that control cell cycle progression . We found that levels of the cyclin-dependent kinase ( CDK ) inhibitor , p27 , were lower in U87MG.DeltaEGFR tumors than in parental U87MG or control U87MG.DK tumors . Consequently , CDK2-cyclin A activity was also elevated , concomitant with the RB protein hyperphosphorylation . In addition , activated phosphatidylinositol 3-kinase ( PI3-K ) and phosphorylated Akt levels were also elevated in the U87MG.DeltaEGFR tumors . U87MG.DeltaEGFR cells failed to arrest in G(1) in response to serum starvation in vitro and while maintaining high levels of PI3-K activity and hyperphosphorylated RB . Treatment of U87MG.DeltaEGFR cells with LY294002 , a PI3-K inhibitor , caused reduced levels of phosphorylated Akt and concomitantly up-regulated levels of p27 . Expression of a kinase dead dominant-negative Akt mutant in the U87MG.DeltaEGFR cells similarly resulted in up-regulation of p27 and down-regulation of tumorigenicity in vivo . These results suggest that the constitutively active DeltaEGFR can enhance cell proliferation in part by down-regulation of p27 through activation of the PI3-K/Akt pathway . This pathway may represent another therapeutic target for treatment of those aggressive glioblastomas expressing DeltaEGFR . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: AMP-activated protein kinase ( AMPK ) has been implicated in anti-proliferative actions in a range of cell systems . Recently , it was observed that Compound C , an inhibitor of AMPK , also reduced the cell viability in human diploid fibroblasts ( HDFs ) . Compound C-induced growth arrest was associated with a decrease in the cell cycle regulatory proteins , such as proliferating cell nuclear antigen , phosphorylated pRB , cyclin-dependent protein kinases ( Cdk 2 and 4 ) , cyclins ( D and E ) , and the Cdk inhibitors ( p21 , p16 , and p27 ) . Therefore , the present study examined the molecular mechanism of the antiproliferative effects of Compound C. Although Compound C inhibited serum-induced phosphorylation of Akt and its substrate , glycogen synthase kinase-3β , it did not affect the Akt activity in vitro . Compound C significantly inhibited the receptor tyrosine phosphorylation and the activity of downstream signaling molecules , such as p85 phosphoinositide 3-kinase , phospholipase C-γ1 , and extracellular signal-regulated kinase 1/2 , induced by platelet-derived growth factor ( PDGF ) but not by epidermal growth factor- and insulin-like growth factor . In vitro growth factor receptor tyrosine kinase activity profiling revealed the IC(50) for PDGF receptor-β ( PDGFRβ ) to be 5.07 μM , whereas the IC(50) for the epidermal growth factor receptor and insulin-like growth factor receptor was ≥100 μM . The inhibitory effect of Compound C on PDGFRβ and Akt was also observed in AMPKα(1)/α(2)-knockout mouse embryonic fibroblasts , indicating that its inhibitory effect is independent of the AMPK activity . The inhibitory effect of Compound C on cell proliferation and PDGFRβ tyrosine phosphorylation was also demonstrated in various PDGFR-expressing cells , including MRC-5 , BEAS-2B , rat aortic vascular smooth muscle cells , and A172 glioblastoma cells . These results indicate that Compound C can be used as a potential antiproliferative agent for PDGF- or PDGFR-associated diseases , such as cancer , atherosclerosis , and fibrosis . OUTPUT: sustaining proliferative signaling INPUT: BACKGROUND Patients with invasive breast ductal carcinoma ( IBDC ) with metastasis have a very poor prognosis . Little is known about the synergistic action of growth and inflammatory factors in IBDC metastases . METHODS The expression of activated extracellular signal-regulated kinase1/2 ( phosphorylated or p-ERK1/2 ) was analyzed by immunohistochemistry in IBDC tissue samples from 80 cases . BT474 IBDC cell migration and invasion were quantified using the Transwell assay . Matrix metalloproteinase ( MMP)-9 expression and activity were analyzed by RT-PCR , Western blotting and zymography . Activator protein ( AP)-1 activity was measured with a luciferase reporter gene assay . The Wilcoxon signed-rank test , Chi-square test , the partition of Chi-square test , independent t-test , and Spearman's method were used for the statistical analysis . RESULTS Phosphorylated ERK1/2 was detected in 58/80 ( 72.5% ) IBDC tissues , and was associated with higher TNM stage and lymph node metastasis , but not patient age or tumor size . Individually , epidermal growth factor ( EGF ) , and interleukin ( IL)-1β activated ERK1/2 , increased cell migration and invasion , MMP-9 expression and activity , AP-1 activation in vitro and the expression of p-ERK1/2 was positively correlated with EGF expression levels , as well as IL-1β , MMP-9 and c-fos in IBDC tissue samples . Co-stimulation with EGF and IL-1β synergistically increased ERK1/2 and AP-1 activation , cell migration and invasion , and MMP-9 expression and activity . Inhibition of ERK1/2 using U0126 or siRNA abolished EGF and/or IL-1β-induced cell migration and invasion in a dose-dependent manner . CONCLUSION Activated ERK1/2 was associated with higher TNM stage and lymph node metastasis in IBDC . Both in vitro and in vivo studies indicated that ERK-1/2 activation may increase the metastatic ability of IBDC cells . Growth and inflammatory factors synergistically induced IBDC cell migration and invasion via ERK1/2 signaling , AP-1 activation and MMP-9 upregulation . OUTPUT: activating invasion and metastasis;sustaining proliferative signaling;tumor promoting inflammation INPUT: PURPOSE Osteosarcoma is an aggressive primary bone cancer characterized by expression of platelet-derived growth factor ( PDGF ) and its cognate receptor . Coexpression of the growth factor and receptor suggests their role in autocrine or paracrine growth mechanisms . It has been reported previously that STI571 has specific activity in inhibiting select tyrosine kinase receptors , including PDGF and c-Kit . Osteosarcomas express low levels of c-Kit but abundant levels of PDGF receptor ( PDGFR ) . EXPERIMENTAL DESIGN To investigate the potential of STI571 as therapy for osteosarcoma , we studied its effects on PDGF-mediated cell growth in vitro and in an in vivo mouse model . RESULTS PDGF acted as a potent mitogen in a dose-dependent manner in two osteosarcoma cell lines . STI571 ( 1.0 micro M ) inhibited both PDGFR-alpha and PDGFR-beta phosphorylation and the downstream phosphorylation targets extracellular signal-regulated kinase and Akt . STI571 also inhibited PDGF-mediated growth and induced apoptosis in vitro as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and terminal deoxynucleotidyl transferase-mediated nick end labeling staining . To study the effect of STI571 alone or in combination with Taxol in an in vivo model , an osteosarcoma cell line ( KRIB ) was transplanted into the tibia of athymic nude mice . Mice were treated with STI571 ( 50 mg/kg p.o. q M-F ) , Taxol ( 8 mg/kg i.p. weekly ) , or STI571 plus Taxol for 6 weeks . There was no significant difference in tumor size between treatment and control mice . Aberrant signaling pathways downstream of the PDGFR in the v-Ki-ras oncogene-transformed KRIB cell line may in part explain this finding . CONCLUSIONS Our data demonstrate that STI571 inhibits PDGF-mediated growth and leads to apoptosis of osteosarcoma cells in vitro by selective inhibition of the PDGFR tyrosine kinase . The effectiveness of STI571 in our studies suggests targeting of PDGFRs as a novel treatment for osteosarcoma . OUTPUT: sustaining proliferative signaling;resisting cell death INPUT: UNLABELLED Activation of beta-catenin , the central effector of the canonical Wnt pathway and a recognized oncogene , has been implicated in hepatocellular carcinoma . We examined N-nitrosodiethylamine ( DEN)-induced tumorigenesis in hepatic beta-catenin conditional knockout mice ( beta-cat KO ) . Male beta-cat KO and age- and sex-matched littermate controls were given a single intraperitoneal DEN injection and followed for 6-12 months for hepatic tumors . Hepatic tumors were characterized for histology , proliferation , apoptosis , oxidative stress , and specific proteins by way of western blot , immunohistochemistry , and coprecipitation studies . For in vivo tumor intervention studies , specific inhibitors were administered intraperitoneally or through drinking water . Intriguingly , beta-cat KO mice showed a paradoxical increase in susceptibility to DEN-induced tumorigenesis . This accelerated tumorigenesis is due to increased injury and inflammation , unrestricted oxidative stress , fibrosis , and compensatory increase in hepatocyte proliferation secondary to platelet-derived growth factor receptor alpha ( PDGFRalpha)/phosphoinositide 3-kinase ( PIK3CA)/Akt activation and c-Myc overexpression . In vitro suppression of beta-catenin expression in hepatoma cells led to enhanced PDGFRalpha expression , which was abrogated in the presence of nuclear factor kappaB ( NF-kappaB ) inhibitor . Daily treatment of 6-month-old DEN-exposed beta-cat KO with PDGFRalpha inhibitor dramatically reduced tumor numbers and size . Inclusion of N-acetyl-L-cysteine , a known antioxidant and NF-kappaB inhibitor , in the drinking water led to complete abolition of tumorigenesis in DEN-exposed beta-cat KO . CONCLUSION Loss of beta-catenin impairs the liver's ability to counteract DEN-induced oxidative stress and enhances tumorigenesis through PDGFRalpha/PIK3CA/Akt signaling . Blockade of PDGFRalpha or oxidative stress dramatically affects beta-catenin-deficient tumorigenesis . Also , hepatoma cells use PDGFRalpha/PIK3CA signaling as an escape mechanism following beta-catenin suppression , and their sequential suppression profoundly impedes tumor proliferation . OUTPUT: tumor promoting inflammation INPUT: OBJECT The intracellular events transducing mitogenic signals from platelet-derived growth factor-beta ( PDGFbeta ) receptor tyrosine kinases are not precisely known . In this study the authors evaluated whether the phosphatidylinositol 3-kinase ( PI3-K)-Akt-p70S6K pathway is expressed in meningiomas , regulates their growth , and transduces mitogenic signals of PDGF-BB . METHODS Nine meningioma tumors obtained in humans were evaluated using Western blot analysis for phosphorylated ( activated ) Akt and phosphorylated p70S6K . Cells cultured from seven of these meningiomas were also screened using Western blot analysis for Akt and for phosphorylated Akt and p70S6K . The authors also evaluated whether PDGF-BB stimulation of meningioma cells was associated with the phosphorylation of Akt and p70S6K known to activate these kinases . In addition , the effects of wortmannin , an inhibitor of P13-K , on proliferation and activation of Akt and p70S6K in meningioma cells stimulated with PDGF-BB were evaluated . Western blots of lysates from meningiomas demonstrated phosphorylated Akt and p70S6K . Treatment with PDGF-BB stimulated phosphorylation of Akt and p70S6K in each meningioma cell culture . Wortmannin ( 500 and 1000 nM ) significantly decreased PDGF-BB stimulation of meningioma cells ( p &lt ; 0.001 ) while it reduced Akt and p70S6K phosphorylation but not mitogen-activated protein kinase/extracellular signal-regulated kinase ( MAPK/ERK ) phosphorylation . CONCLUSIONS These findings indicate that Akt and p70S6K are constitutively expressed and activated in meningioma cells and that the PI3-K-Akt-p70S6K pathway may participate in transduction of mitogenic signals in meningiomas independent of the Raf-1-MEK-1-MAPK/ERK cascade . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot15

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND/AIMS The use of chemotherapy in hepatocellular carcinoma is still controversial . The aim of this study was to investigate whether the combined use of epirubicin and progesterone has a synergistic effect on cell proliferation and apoptosis in HepG2 cells . METHODOLOGY Different concentrations of epirubicin ( 0.1 microg/ml , 0.25 microg/ml and 0.5microg/ml ) or progesterone ( 12.5 microM , 25 microM and 50 microM ) were added to HepG2 cells either alone or in combinations consisting of different concentrations of the two . Their effects on HepG2 cells were studied by ( 1 ) XTT assay for analysis of cell proliferation , ( 2 ) 3H-Thymidine incorporation for DNA synthesis , ( 3 ) annexin V-FITC/ propidium iodide ( PI ) flowcytometery for cell apoptosis , ( 4 ) flowcytometry for cell cycle distributions , and ( 5 ) reverse transcription-polymerase chain reaction for expression of cell cycle modulator , cyclin D1 . RESULTS 50 microM progesterone increased both the cytotoxic and apoptotic effects of 0.1 microg/ml epirubicin on HepG2 cells at 48 hr culture due to 50 microM progesterone accumulated cells in S phase of the cell cycle and subsequently reduced cyclin D1 expression . These effects on HepG2 cells induced by this combination were comparable to those induced by 0.5 microg/ml epirubicin alone . CONCLUSIONS In vitro , progesterone can increase the cytotoxicity and apoptosis induced by epirubicin on HepG2 cells . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Recent studies indicate that cyclooxygenase-2 ( COX-2 ) is overexpressed in pancreatic adenocarcinoma and may play a critical role in this rapidly progressing form of cancer . A human pancreatic adenocarcinoma cell line , Mia PaCa-2 , was incubated for 18 hours with 5 micromol/L of rofecoxib ( Vioxx ) , a selective COX-2 inhibitor . Total RNA was isolated and gene expression analyzed by DNA microarray chips . In a separate experiment , athymic mice were orthotopically injected with 7.5 x 10(5) Mia PaCa-2 cells through a minilaparotomy . After 1 month , laparotomy was repeated to measure tumor size , and mice were randomized to receive reformulated rodent chow containing either 12.5 mg/kg/day of rofecoxib or no drug for 21 days . Tumor growth was assessed by comparing volume before and after treatment . In vitro , rofecoxib decreased gene expression of cyclin D1/PRAD1 , a key component of cell cycle progression , while increasing expression of several cell cycle arrest genes , including p21/WAF1 , p33/ING , GADD34 , and GADD45 ( P &lt ; 0.05 ) . In vivo , tumor growth was significantly reduced in treated vs. control mice ( P &lt ; 0.05 ) . No systemic toxicity was observed in mice receiving rofecoxib . These data suggest that rofecoxib slows the growth of human pancreatic cancer through changes in gene expression that favor cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Certain hexavalent chromium ( Cr(VI) ) compounds are well established occupational respiratory tract carcinogens . However , despite extensive studies , the cellular and molecular mechanisms underlying Cr(VI)-induced lung cancer remain poorly understood . In fact , the models used were often suboptimal and yielded conflicting results that were heavily dependent upon the system and experimental conditions employed . Here , we investigated the effects of chronic subcytotoxic and mildly cytotoxic ( 0.1-2 microM ) Cr(VI) exposures on cultures of human bronchial epithelial cells , the main targets of Cr(VI) carcinogenicity . Our studies with the nontumorigenic BEAS-2B cell line suggest that relatively short exposures ( h ) to sublethal Cr(VI) doses ( 0.1-1 microM ) may render these cells less sensitive to contact inhibition . We have also observed a reduced sensitivity to Cr(VI)-induced apoptosis shortly after the beginning of exposure to a mildly cytotoxic Cr(VI) dose ( 2 microM ) . Further studies are needed to determine whether these two phenotypes are involved in the Cr(VI)-induced carcinogenic process . Additionally , evidence gathered in this study strongly points to a Cr(VI) interference with cell adhesion to the substratum and with cell-cell interactions . Finally , by chronically exposing BEAS-2B cells to mildly cytotoxic Cr(VI) doses ( 1 and 2 microM ) , we were able to induce changes in cell morphology and pattern of growth characteristic of an early phase of pre-malignant progression . OUTPUT: evading growth suppressors;resisting cell death INPUT: OBJECTIVE To study the effects of genistein on the proliferation , apoptosis induction and expression of related gene proteins of human colon cancer cells in vitro and in vivo , and its mechanisms of action . METHODS MTT colorimetric assay was used to detect the effects of genistein on the proliferation of human colon adenocarcinoma SW480 cells . Light and transmission electron microscopy were used to study the histological and ultrastructural changes . Flow cytometry was used to determine the effects of genistein on cell cycle and apoptosis . Flow cytometry and immunohistochemistry were used to determine the effects of genistein on apoptosis induction and expression of related gene proteins of colon cancer cells . RESULTS The MTT colorimetric assay showed that genistein inhibited the proliferation of SW480 cells in a dose-dependent and time-dependent manner , and the highest inhibition rate was 60.2% after 80 microg/ml genistein treatment for 72 h . The light microscopy revealed that many genistein-treated cancer cells were shrunken , disrupted , or showing cytoplasmic vacuolization . The electron microscopic examination showed cell shrinkage , nuclear fragmentation and pronounced chromatin condensation , sometimes formed crescent chromatin condensation attached to the nuclear membrane . The results of flow cytometry showed that : after SW480 cells were treated with 0 , 20 , 40 , 80 microg/ml genistein for 48 h , the FI values of PCNA were 1.49 +/- 0.02 , 1.28 +/- 0.04 , 1.14 +/- 0.03 , and 0.93 +/- 0.08 ; the FI values of VEGF were 1.75 +/- 0.02 , 1.34 +/- 0.06 , 1.32 +/- 0.04 , and 1.23 +/- 0.04 ; the fluorescence index ( FI ) values of p21 were 1.26 +/- 0.05 , 1.36 +/- 0.06 , 1.61 +/- 0.03 , and 1.73 +/- 0.03 , respectively . There were statistically significant differences between the control group and each treatment group ( P &lt ; 0.05 or P &lt ; 0.01 ) . The scores of immunohistochemical staining of PCNA and VEGF proteins were decreased , while p21 increased . There were statistically significant differences between the control group and each treatment group ( P &lt ; 0.05 or P &lt ; 0.01 ) . CONCLUSION Genistein can inhibit the growth of colon cancer cells via apoptosis induction and cell cycle arrest at G(2)/M phase . The anti-tumor mechanisms of genistein may be related with the down-regulation of expression of VEGF and PCNA , and up-regulation of the expression of p21 . OUTPUT: resisting cell death;evading growth suppressors;sustaining proliferative signaling;inducing angiogenesis INPUT: In human colorectal adenomas or polyps , cyclooxygenase-2 is expressed predominantly by stromal ( or interstitial ) macrophages . Therefore , we tested the hypothesis that macrophage cyclooxygenase-2 has paracrine pro-tumorigenic activity using in vitro models of macrophage-epithelial cell interactions . We report that macrophages can promote tumorigenic progression of intestinal epithelial cells ( evidenced by decreased cell-cell contact inhibition , increased proliferation and apoptosis , gain of anchorage-independent growth capability , decreased membranous E-cadherin expression , up-regulation of cyclooxygenase-2 expression , down-regulation of transforming growth factor-beta type II receptor expression and resistance to the anti-proliferative activity of transforming growth factor-beta(1) ) in a paracrine , cyclooxygenase-2-dependent manner . Pharmacologically relevant concentrations ( 1-2 microM ) of a selective cyclooxygenase-2 inhibitor had no detectable , direct effect on intestinal epithelial cells but inhibited the macrophage-epithelial cell signal mediating tumorigenic progression . Cyclooxygenase-2-mediated stromal-epithelial cell signalling during the early stages of intestinal tumorigenesis provides a novel target for chemoprevention of colorectal cancer ( and other gastro-intestinal epithelial malignancies , which arise on a background of chronic inflammation , such as gastric cancer ) and may explain the discrepancy between the concentrations of cyclooxygenase inhibitors required to produce anti-neoplastic effects in vitro and in vivo . OUTPUT:

evading growth suppressors;resisting cell death;sustaining proliferative signaling;tumor promoting inflammation

HoC_dynamic_5_shot16

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Telomerase is a ribonucleoprotein consisting of a catalytic subunit , the telomerase reverse transcriptase , TERT , and an integrally associated RNA , TR , which contains a template for the synthesis of short repetitive G-rich DNA sequences at the ends of telomeres . Telomerase can repetitively reverse transcribe its short RNA template , acting processively to add multiple telomeric repeats onto the same DNA substrate . The contribution of enzyme processivity to telomere length regulation in human cells is not well characterized . In cancer cells , under homeostatic telomere length-maintenance conditions , telomerase acts processively , while under nonequilibrium conditions , telomerase acts distributively on the shortest telomeres . To investigate the role of increased telomerase processivity on telomere length regulation in human cells with limited lifespan that are dependent on human TERT ( hTERT ) for lifespan extension and immortalization , we mutated the leucine at position 866 in the reverse transcriptase C motif of hTERT to a tyrosine ( L866Y ) , which is the amino acid found at a similar position in HIV-1 reverse transcriptase . We report that , similar to the previously reported ' gain of function ' Tetrahymena telomerase mutant ( L813Y ) , the human telomerase variant displays increased processivity. hTERT-L866Y , like wild-type hTERT can immortalize and extend the lifespan of limited lifespan cells . Moreover , hTERT-L866Y expressing cells display heterogenous telomere lengths , telomere elongation , multiple telomeric signals indicative of fragile sites and replicative stress , and an increase in short telomeres , which is accompanied by telomere trimming events . Our results suggest that telomere length and homeostasis in human cells may be regulated by telomerase enzyme processivity . OUTPUT: enabling replicative immortality INPUT: Telomerase is a ribonucleoprotein enzyme that functions to maintain telomeres , the terminal DNA that protects chromosomal integrity , regulating cellular replicative life span . Telomerase is not expressed in most normal human somatic cells but is active in stabilizing telomeres of certain self-renewing cell populations and most malignant cells , making the enzyme an appealing target for anticancer therapy . We describe here a novel cross-species approach to telomerase inhibition . Ectopic expression of the human telomerase catalytic reverse transcriptase component in murine cells inhibited endogenous murine telomerase activity . Using this approach , telomerase inhibition in immortal murine fibroblasts resulted in critical telomere shortening , leading to slowed proliferation , abnormal morphology , altered cell cycle , and telomere dysfunction with cytogenetic instability , followed by apoptotic cell death . Subpopulations of two telomerase-inhibited clones escaped widespread apoptosis , showing proliferative recovery in culture despite persistently inhibited telomerase activity with progressive telomere shortening and dysfunction . This study , by targeting immortal murine cells for telomerase inhibition , demonstrates the importance of telomerase to murine cell immortalization and telomere maintenance . Moreover , the murine model used here should prove useful in further evaluating telomerase inhibition as an anticancer therapy . OUTPUT: enabling replicative immortality;resisting cell death INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: The vast majority of breast cancers are carcinomas that arise from mammary epithelial cells ( MECs ) . One of the key early events in tumorigenic transformation is the ability of cells to overcome replicative senescence . However , the precise genetic changes that are responsible for this event in MECs is largely unknown . Here , we report that Bmi-1 , originally identified as a c-Myc cooperating oncoprotein , can bypass senescence , extend the replicative life span , and immortalize MECs . Furthermore , Bmi-1 was overexpressed in immortal MECs and several breast cancer cell lines . Overexpression of Bmi-1 in MECs led to activation of human telomerase reverse transcriptase ( hTERT ) transcription and induction of telomerase activity . Telomerase induction by Bmi-1 was an early event in the extension of the replicative life span and immortalization . Bmi-1 was not overexpressed in hTERT-immortalized MECs , suggesting that Bmi-1 functions upstream of hTERT . Although , c-Myc has been reported to induce telomerase in MECs , Bmi-1 appeared to act independently of c-Myc binding sequences in the hTERT promoter . Deletion analysis of the Bmi-1 protein suggested that the RING finger , as well as a conserved helix-turn-helix-turn domain , were required for its ability to induce telomerase and immortalize MECs . These data suggest that Bmi-1 regulates telomerase expression in MECs and plays a role in the development of human breast cancer . OUTPUT: enabling replicative immortality INPUT: BACKGROUND The most deadly form of cancer is not lung or colon , breast or prostate ; it is any cancer that has become metastatic . Mortality due to metastatic melanoma , one of the most aggressive and deadly cancers , has increased steadily over the last several decades . Unfortunately , the arsenal of chemotherapeutic agents available today is most often unsuccessful at extending and improving the life expectancy of afflicted individuals . We sought to identify an effective and nontoxic agent against metastatic melanoma . METHODOLOGY/PRINCIPAL FINDINGS We chose to study Cloudman S-91 mouse melanoma cells ( sub-clone M3 , CCL53.1 ) because these cells are highly aggressive and metastatic , representing one of the deadliest types of cancer . Melanoma cells also had an experimental advantage because their morphology , which is easily monitored , relates to the physiology of metastatic cells and normal melanocytes . We chose to test methyl sulfone as a chemotherapeutic agent for two reasons . Because of its chemical structure , we speculated a potential anti-cancer activity by targeting microtubules . Equally important , methyl sulfone has a well-established safety profile in humans . Surprisingly , we found that malignant melanoma cells exposed to methyl sulfone demonstrated the loss of phenotypes characteristic of malignant cells , and the reemergence of phenotypes characteristic of healthy melanocytes . Briefly , over time methyl sulfone induced contact inhibition , loss of ability to migrate through an extracellular matrix , loss of anchorage-independent growth , proper wound healing followed by contact inhibition , irreversible senescence followed by arborization with melanosomes in arbors as seen in normal melanocytes . CONCLUSIONS/SIGNIFICANCE Methyl sulfone may have clinical potential as a non-toxic agent effective against metastatic melanoma . Additionally , methyl sulfone has promise as a tool to explore molecular mechanisms of metastatic transformation as well as fundamental processes such as cell migration , contact inhibition , wound healing and cellular senescence . OUTPUT: enabling replicative immortality;evading growth suppressors;activating invasion and metastasis INPUT: A hallmark of cancer cells is the ability to proliferate indefinitely . This acquisition of an immortal lifespan usually requires the activation of telomerase , the enzyme that elongates telomeres . Human telomerase is minimally composed of the reverse transcriptase subunit hTERT , and the RNA subunit hTR . While hTR is ubiquitously expressed in human cells , the hTERT subunit is generally transcriptionally repressed in most normal somatic cells , but is illegitimately activated to restore telomerase activity in cancer cells . Indeed , in the thousands of different human tumours assayed , 85% were scored positive for telomerase activity . However , the levels of telomerase activity detected in tumour samples can vary substantially and even some normal somatic cells have been found to have low levels of enzyme activity . As the functional significance of low levels of telomerase activity is unclear , we investigated whether there is a minimum level of telomerase activity required for tumourigenesis . Using mutants of hTERT that induce varying levels of telomerase activity , we show that there does indeed exist a threshold of activity required for the processes of immortalization , transformation and tumourigenesis . Thus , low levels of activity detected in certain somatic cells would not be expected to contribute to tumourigenesis , nor does the mere detection of telomerase in cancer cells necessarily signify an immortal lifespan . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot17

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Engineered zinc-finger nucleases ( ZFN ) represented an innovative method for the genome manipulation in vertebrates . ZFN introduced targeted DNA double strand breaks ( DSB ) and initiated non-homologous end joining ( NHEJ ) after pronuclear or cytoplasmatic microinjection into zygotes . Resulting frame shift mutations led to functional gene ablations in zebra fish , mice , pigs and also in laboratory rats . Therefore , we targeted the rat Rag1 gene essential for the V(D)J recombination within the immunoglobulin production process and for the differentiation of mature B and T lymphocytes to generate an immunodeficient rat model in the LEW/Ztm strain . RESULTS After microinjection of Rag1 specific ZFN mRNAs in 623 zygotes of inbred LEW/Ztm rats 59 offspring were born from which one carried a 4 bp deletion . This frame shift mutation led to a premature stop codon and a subsequently truncated Rag1 protein confirmed by the loss of the full-length protein in Western Blot analysis . Truncation of the Rag1 protein was characterized by the complete depletion of mature B cells . The remaining T cell population contained mature CD4+/CD3+/TCRαβ+ as well as CD8+/CD3+/TCRαβ+ positive lymphocytes accompanied by a compensatory increase of natural killer cells in the peripheral blood . Reduction of T cell development in Rag1 mutant rats was associated with a hypoplastic thymus that lacked follicular structures . Histological evaluation also revealed the near-complete absence of lymphocytes in spleen and lymph nodes in the immunodeficient Rag1 mutant rat . CONCLUSION The Rag1 mutant rat will serve as an important model for transplantation studies . Furthermore , it may be used as a model for reconstitution experiments related to the immune system , particularly with respect to different populations of human lymphocytes , natural killer cells and autoimmune phenomena . OUTPUT: genomic instability and mutation;avoiding immune destruction INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: Helicobacter hepaticus-infected Rag2(-/-) mice emulate many aspects of human inflammatory bowel disease , including the development of colitis and colon cancer . To elucidate mechanisms of inflammation-induced carcinogenesis , we undertook a comprehensive analysis of histopathology , molecular damage , and gene expression changes during disease progression in these mice . Infected mice developed severe colitis and hepatitis by 10wk post-infection , progressing into colon carcinoma by 20wk post-infection , with pronounced pathology in the cecum and proximal colon marked by infiltration of neutrophils and macrophages . Transcriptional profiling revealed decreased expression of DNA repair and oxidative stress response genes in colon , but not in liver . Mass spectrometric analysis revealed higher levels of DNA and RNA damage products in liver compared to colon and infection-induced increases in 5-chlorocytosine in DNA and RNA and hypoxanthine in DNA . Paradoxically , infection was associated with decreased levels of DNA etheno adducts . Levels of nucleic acid damage from the same chemical class were strongly correlated in both liver and colon . The results support a model of inflammation-mediated carcinogenesis involving infiltration of phagocytes and generation of reactive species that cause local molecular damage leading to cell dysfunction , mutation , and cell death . There are strong correlations among histopathology , phagocyte infiltration , and damage chemistry that suggest a major role for neutrophils in inflammation-associated cancer progression . Further , paradoxical changes in nucleic acid damage were observed in tissue- and chemistry-specific patterns . The results also reveal features of cell stress response that point to microbial pathophysiology and mechanisms of cell senescence as important mechanistic links to cancer . OUTPUT: tumor promoting inflammation;genomic instability and mutation;resisting cell death INPUT: BACKGROUND &amp ; OBJECTIVE Altered cell adhesion has a critical role in the development of epithelial cancers . E-cadherin act on the maintenance of cell-cell adhesion and its function was thought to be regulated by associated cytoplasmic proteins , such as alpha-catenin and beta-catenin . This study was designed to examine the expression of beta-catenin in gastric carcinoma and to determine the relationship between tumor characteristics and survival . METHODS Immunohistochemical staining of beta-catenin and E-cadherin was performed in 148 patients with gastric carcinoma . RESULTS Abnormal expression of beta-catenin and E-cadherin was performed in 148 patients with gastric carcinoma . RESULTS Abnormal expression of beta-catenin and E-cadherin was demonstrated in 43.2% and 44.6% of tumors respectively . Up to 63% of tumors stained abnormally for one or two components of the cadherin-catenin complex ( beta-catenin , E-cadherin ) . Abnormal beta-catenin and E-cadherin staining occurred more frequently in poor differentiated tumor than in good differentiated tumors ( P &lt ; 0.005 , respectively ) . There was a significant correlation between abnormal beta-catenin expression and depth of invasion(P &lt ; 0.025 ) . Moreover , abnormal beta-catenin expression was more frequent in tumors with positive lymph node metastasis ( 45/84 , 53.6% ) and distance metastasis(21/31 , 67.7% ) than in tumors without lymph node metastasis ( 19/64 , 29.7% ) ( P &lt ; 0.005 ) and distance metastasis ( 43/117 , 36.8% ) ( P &lt ; 0.005 ) . A survival advantage was noted in tumors retaining normal membranous expression of beta-catenin , independent of type , grade , or stage ( P &lt ; 0.005 ) . CONCLUSIONS Abnormal expression of the E-cadherin-catenin complex occurs frequently in gastric carcinoma . The close correlation with poor survival suggests that abnormal beta-catenin may be a useful prognostic marker . OUTPUT: activating invasion and metastasis INPUT: Breast cancer is the malignant neoplasia with the highest incidence in women worldwide . Chronic oxidative stress and inflammation have been indicated as major mediators during carcinogenesis and cancer progression . Human studies have not considered the complexity of tumor biology during the stages of cancer advance , limiting their clinical application . The purpose of this study was to characterize systemic oxidative stress and immune response parameters in early ( ED ; TNM I and II ) and advanced disease ( AD ; TNM III and IV ) of patients diagnosed with infiltrative ductal carcinoma breast cancer . Oxidative stress parameters were evaluated by plasmatic lipoperoxidation , carbonyl content , thiobarbituric reactive substances ( TBARS ) , nitric oxide levels ( NO ) , total radical antioxidant parameter ( TRAP ) , superoxide dismutase , and catalase activities and GSH levels . Immune evaluation was determined by TNF-α , IL-1β , IL-12 , and IL-10 levels and leukocytes oxidative burst evaluation by chemiluminescence . Tissue damage analysis included heart ( total CK and CKMB ) , liver ( AST , ALT , GGT ) , and renal ( creatinine , urea , and uric acid ) plasmatic markers . C-reactive protein ( CRP ) and iron metabolism were also evaluated . Analysis of the results verified different oxidative stress statuses occur at distinct cancer stages . ED was characterized by reduction in catalase , 8-isoprostanes , and GSH levels , with enhanced lipid peroxidation and TBARS levels . AD exhibited more pronounced oxidative status , with reduction in catalase activity and TRAP , intense lipid peroxidation and high levels of NO , TBARs , and carbonyl content . ED patients presented a Th2 immune pattern , while AD exhibited Th1 status . CRP levels and ferritin were increased in both stages of disease . Leukocytes burst impairment was observed in both the groups . Plasma iron levels were significantly elevated in AD . The data obtained indicated that oxidative stress enhancement and immune response impairment may be necessary to ensure cancer progression to advanced stages and may result from both host and tumor inflammatory mediators . OUTPUT: tumor promoting inflammation INPUT: Environmental carcinogen exposure may play an important role in the incidence of cancer in children . In addition to environmental pollutants , maternal smoking during pregnancy may be a contributing factor . Major carcinogenic components of cigarette smoke and other combustion by-products in the environment include polycyclic aromatic hydrocarbons ( PAH ) . Mouse offspring exposed during midpregnancy to the PAH , benzo[a]pyrene ( B[a]P ) , show significant deficiencies in their immune functions , observed in late gestation which persist for at least 18 months . Tumor incidences in these progeny are 8 to 10-fold higher than in controls . We have demonstrated a significant reduction in thymocytes ( CD4+ CD8+ , CD4+ CD8+ Vbeta8+ , CD4+ CD8+ Vgamma2+ ) from newborn and splenocytes ( CD4+ CD8+ ) from 1-week-old mouse progeny exposed to B[a]P in utero . To investigate possible causes of the observed T cell reduction , we analyzed the thymocytes and splenocytes from progeny and maternal tissues for the presence of B[a]P-DNA adducts . Adducts were detected in maternal , placental and offspring lymphoid tissues at day 19 of gestation , at birth and 1-wk after birth . The presence of B[a]P-DNA adducts in immature T cells may , in part , explain the previously observed T cell immunosuppression and tumor susceptibility in mice exposed to B[a]P in utero . The effects of DNA lesions on progeny T cells may include interference with normal T-cell development . These results provide a possible explanation for the relationship between maternal smoking during pregnancy and childhood carcinogenesis . OUTPUT:

avoiding immune destruction;genomic instability and mutation

HoC_dynamic_5_shot18

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Epidermal growth factor receptor-tyrosine kinase inhibitors ( EGFR-TKIs ) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor ( EGFR ) gene . On the other hand , some lung cancer patients with wild type EGFR also respond to EGFR-TKIs , suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells . However , the effect of EGFR-TKIs on host microenvironments is largely unknown . A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells . This model was used to investigate the therapeutic efficacy of erlotinib , an EGFR-TKI , on multiple organ metastases induced by human small cell lung cancer cells ( SBC-5 cells ) that did not express EGFR . Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro , it significantly suppressed bone and lung metastases in vivo , but not liver metastases . An immunohistochemical analysis revealed that , erlotinib significantly suppressed the number of osteoclasts in bone metastases , whereas no difference was seen in microvessel density . Moreover , erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line ( MC3T3-E1 cells ) . These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells . OUTPUT: activating invasion and metastasis;inducing angiogenesis;sustaining proliferative signaling INPUT: The UT-7 cell line was established from a patient with a megakaryoblastic leukemia ( Komatsu et al , Cancer Res 51 : 341 , 1991 ) . Its proliferation is strictly dependent on the presence of hematopoietic growth factors including erythropoietin ( Epo ) , granulocyte-macrophage colony-stimulating factor ( GM-CSF ) , and interleukin-3 ( IL-3 ) . We investigated the differentiation capacities of this cell line under the action of several growth factors , using immunomarkers , flow cytometry , and ultrastructural techniques . In the presence of GM-CSF and IL-3 , eosinophil and basophil promyelocytes were detected , as well as a few cells with erythroid and megakaryocytic ( MK ) differentiation features . In contrast , Epo induced a marked erythroid differentiation with an increase of glycophorin A expression , accompanied by a few hemoglobinized cells . Differentiation induced by the growth factors took 24 to 48 hours to begin , and increased with cell passages to a plateau at 2 weeks of culture . However , this was not only due to a cell selection because the differential effects of Epo and GM-CSF were observed from a single cell clone and the phenotype could be reversed by opposite growth factors , even after a long period of culture . We subsequently investigated the phenotype of UT-7 in the presence of combinations of Epo , IL-3 , and GM-CSF , and showed that GM-CSF and IL-3 act predominantly over Epo . This effect was mediated by a rapid downmodulation of Epo receptors by GM-CSF at messenger RNA and binding sites levels , without a change in receptor affinities . On the other hand , Epo had no effect on number and affinity of GM-CSF receptors . This study shows that UT-7 is a growth factor-dependent pluripotent cell line in which commitment may be directed by a hierarchical action of growth factors through an early and rapid transmodulation of growth factor receptors . OUTPUT: sustaining proliferative signaling INPUT: This study aimed to analyze the role of endothelial progenitor cell ( EPC)-derived angiogenic factors and chemokines in the multistep process driving angiogenesis with a focus on the recently discovered macrophage migration inhibitory factor ( MIF)/chemokine receptor axis . Primary murine and murine embryonic EPCs ( eEPCs ) were analyzed for the expression of angiogenic/chemokines and components of the MIF/CXC chemokine receptor axis , focusing on the influence of hypoxic versus normoxic stimulation . Hypoxia induced an upregulation of CXCR2 and CXCR4 but not CD74 on EPCs and triggered the secretion of CXCL12 , CXCL1 , MIF , and vascular endothelial growth factor ( VEGF ) . These factors stimulated the transmigration activity and adhesive capacity of EPCs , with MIF and VEGF exhibiting the strongest effects under hypoxia . MIF- , VEGF- , CXCL12- , and CXCL1-stimulated EPCs enhanced tube formation , with MIF and VEGF exhibiting again the strongest effect following hypoxia . Tube formation following in vivo implantation utilizing angiogenic factor-loaded Matrigel plugs was only promoted by VEGF . Coloading of plugs with eEPCs led to enhanced tube formation only by CXCL12 , whereas MIF was the only factor which induced differentiation towards an endothelial and smooth muscle cell ( SMC ) phenotype , indicating an angiogenic and differentiation capacity in vivo . Surprisingly , CXCL12 , a chemoattractant for smooth muscle progenitor cells , inhibited SMC differentiation . We have identified a role for EPC-derived proangiogenic MIF , VEGF and MIF receptors in EPC recruitment following hypoxia , EPC differentiation and subsequent tube and vessel formation , whereas CXCL12 , a mediator of early EPC recruitment , does not contribute to the remodeling process . By discerning the contributions of key angiogenic chemokines and EPCs , these findings offer valuable mechanistic insight into mouse models of angiogenesis and help to define the intricate interplay between EPC-derived angiogenic cargo factors , EPCs , and the angiogenic target tissue . OUTPUT: inducing angiogenesis INPUT: Recent studies have demonstrated that n-3 polyunsaturated fatty acids such as eicosapentaenoic acid ( EPA ) are able to suppress cell proliferation and inhibit tumor growth . The objective of our study was to investigate the influence of a high dose EPA on the development of the tumor phenotype in ataxia-telangiectasia mutated ( Atm)-deficient mice , a genetic cancer model that is associated with increased levels of oxidative stress . We analyzed toxicity , proliferation , cell-cycle progression , and apoptosis of EPA in vitro and latency to tumorigenesis in vivo . Because of the impact of reactive oxygen species ( ROS ) on the tumor incidence in ataxia telangiectasia ( AT ) , we further analyzed the effect of EPA on the generation of ROS and oxidative DNA damage ( ODD ) . EPA effectively inhibited proliferation , altered cell-cycle progression , and induced apoptosis of tumor cells ( AT-4 ) . EPA showed no effect on the latency to tumorigenesis in Atm-deficient mice . EPA treatment was accompanied by a significant increase of ROS and ODD . Our results demonstrate the antiproliferative effect of EPA on tumor cells by alteration of cell-cycle progression and induction of apoptosis in vitro . On the other hand , EPA treatment of Atm-deficient mice led to the formation of ROS and accumulation of ODD that might have abrogated the anticarcinogenic effect caused by EPA . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: Estrogen receptor ( ER ) and NF-κB are transcription factors with profound effects on breast cancer cell proliferation and survival . While many studies demonstrate that ER and NF-κB can repress each other , we previously identified a gene signature that is synergistically upregulated by these two factors in more aggressive luminal B breast tumors . Herein , we examine a novel mechanism of cross talk between ER and NF-κB that results in the upregulation of the antiapoptotic gene BIRC3 ( also known as cIAP2 ) . We demonstrate that NF-κB , acting through two response elements , is required for ER recruitment to an adjacent estrogen response element ( ERE ) in the BIRC3 promoter . This effect is accompanied by a major increase in NF-κB-dependent histone acetylation around the ERE . Interestingly , CBP , a histone acetyltransferase previously implicated in repressive interactions between ER and NF-κB , plays a permissive role by promoting histone acetylation and ER recruitment , as well as enhanced expression of BIRC3 . These findings suggest a new gene regulatory mechanism by which inflammation and NF-κB activation can influence ER recruitment to inherently inactive ER binding sites . This fine-tuning mechanism may explain how two factors that generally repress each other's activity may work together on certain genes to promote breast cancer cell survival and tumor progression . OUTPUT: sustaining proliferative signaling;tumor promoting inflammation INPUT: Binding of erythropoietin ( EPO ) to its receptor ( EPOR ) on erythroid cells induces the activation of numerous signal transduction pathways , including the mitogen-activated protein kinase Jun-N-terminal kinase ( JNK ) . In an effort to understand the regulation of EPO-induced proliferation and JNK activation , we have examined the role of potential autocrine factors in the proliferation of the murine erythroleukemia cell line HCD57 . We report here that treatment of these cells with EPO induced the expression and secretion of tumor necrosis factor alpha ( TNF-alpha ) . EPO-dependent proliferation was reduced by the addition of neutralizing antibodies to TNF-alpha , and exogenously added TNF-alpha induced proliferation of HCD57 cells . EPO also could induce TNF-alpha expression in BAF3 and DA3 myeloid cells ectopically expressing EPOR . Addition of TNF-alpha activated JNK in HCD57 cells , and the activity of JNK was partially inhibited by addition of a TNF-alpha neutralizing antibody . Primary human and murine erythroid progenitors expressed TNF-alpha in either an EPO-dependent or constitutive manner . However , TNF-alpha had an inhibitory effect on both immature primary human and murine cells , suggestive that the proliferative effects of TNF-alpha may be limited to erythroleukemic cells . This study suggests a novel role for autocrine TNF-alpha expression in the proliferation of erythroleukemia cells that is distinct from the effect of TNF-alpha in normal erythropoiesis . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot19

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Previous epidemiologic observational and experimental studies investigated the potential of antioxidant micronutrients to modulate cancer risk , but these studies produced inconsistent results . In this pilot , randomized , double-blind , placebo-controlled clinical trial ( n = 47 ) , we assessed the effects of an antioxidant micronutrient combination ( 800 mg dl-alpha-tocopherol acetate , 24 mg beta-carotene , 1.0 g vitamin C , 200 microg l-selenomethionine , 7.2 mg riboflavin , 80 mg niacin , 60 mg zinc , 5 mg manganese ) given daily over 4 months on oxidative and inflammatory biomarkers in patients with a history of sporadic colorectal adenoma . Plasma tumor necrosis factor-alpha ( TNF-alpha ) , interleukin-6 , and F2-isoprostane concentrations were measured using ELISAs , and cystine ( CySS ) was measured using high-performance liquid chromatography . Plasma TNF-alpha concentration decreased in the active treatment group by 37% relative to the placebo group ( P = 0.002 ) , and CySS decreased by 19% ( P = 0.03 ) ; however , interleukin-6 and F2-isoprostane concentrations decreased in antioxidant-treated nonsmokers but increased in smokers , although these findings were not statistically significant . The decreases of TNF-alpha and CySS were more pronounced in nonsmokers . These data suggest that ( a ) an antioxidant micronutrient cocktail can modulate biomarkers of oxidative stress and inflammation in humans and ( b ) the effects of antioxidant micronutrient supplementation on biomarkers of inflammation and oxidative stress may differ according to smoking status . OUTPUT: tumor promoting inflammation INPUT: UNLABELLED Activation of beta-catenin , the central effector of the canonical Wnt pathway and a recognized oncogene , has been implicated in hepatocellular carcinoma . We examined N-nitrosodiethylamine ( DEN)-induced tumorigenesis in hepatic beta-catenin conditional knockout mice ( beta-cat KO ) . Male beta-cat KO and age- and sex-matched littermate controls were given a single intraperitoneal DEN injection and followed for 6-12 months for hepatic tumors . Hepatic tumors were characterized for histology , proliferation , apoptosis , oxidative stress , and specific proteins by way of western blot , immunohistochemistry , and coprecipitation studies . For in vivo tumor intervention studies , specific inhibitors were administered intraperitoneally or through drinking water . Intriguingly , beta-cat KO mice showed a paradoxical increase in susceptibility to DEN-induced tumorigenesis . This accelerated tumorigenesis is due to increased injury and inflammation , unrestricted oxidative stress , fibrosis , and compensatory increase in hepatocyte proliferation secondary to platelet-derived growth factor receptor alpha ( PDGFRalpha)/phosphoinositide 3-kinase ( PIK3CA)/Akt activation and c-Myc overexpression . In vitro suppression of beta-catenin expression in hepatoma cells led to enhanced PDGFRalpha expression , which was abrogated in the presence of nuclear factor kappaB ( NF-kappaB ) inhibitor . Daily treatment of 6-month-old DEN-exposed beta-cat KO with PDGFRalpha inhibitor dramatically reduced tumor numbers and size . Inclusion of N-acetyl-L-cysteine , a known antioxidant and NF-kappaB inhibitor , in the drinking water led to complete abolition of tumorigenesis in DEN-exposed beta-cat KO . CONCLUSION Loss of beta-catenin impairs the liver's ability to counteract DEN-induced oxidative stress and enhances tumorigenesis through PDGFRalpha/PIK3CA/Akt signaling . Blockade of PDGFRalpha or oxidative stress dramatically affects beta-catenin-deficient tumorigenesis . Also , hepatoma cells use PDGFRalpha/PIK3CA signaling as an escape mechanism following beta-catenin suppression , and their sequential suppression profoundly impedes tumor proliferation . OUTPUT: tumor promoting inflammation INPUT: Common use of antimutagens and anticarcinogens in everyday life is an effective measure for preventing human cancer and genetic diseases . Antioxidant properties of tea have vast potential as protective agents against diverse toxic effects . The present study was aimed to evaluate the role of aqueous clonal tea extracts ( green tea , oolong tea and black tea ) in modulating the genotoxic damage induced by cyclophosphamide ( CP ) , a commonly used chemotherapeutic drug and a well-known mutagen and clastogen . All the three tea extracts at 1 and 2% concentration did not increase the frequency of micronucleated polychromatic erythrocytes ( MPE ) in bone marrow cells of mice when administered individually . The tea extracts decreased the micronuclei ( MN ) induced by CP . Therefore , regular intake of tea may improve the antioxidant status in in vivo and thereby reduce the risk of cancer and coronary heart disease . OUTPUT: genomic instability and mutation INPUT: The recent approval of a prostate cancer vaccine has renewed hope for anticancer immunotherapies . However , the immunosuppressive tumor microenvironment may limit the effectiveness of current immunotherapies . Antiangiogenic agents have the potential to modulate the tumor microenvironment and improve immunotherapy , but they often are used at high doses in the clinic to prune tumor vessels and paradoxically may compromise various therapies . Here , we demonstrate that targeting tumor vasculature with lower vascular-normalizing doses , but not high antivascular/antiangiogenic doses , of an anti-VEGF receptor 2 ( VEGFR2 ) antibody results in a more homogeneous distribution of functional tumor vessels . Furthermore , lower doses are superior to the high doses in polarizing tumor-associated macrophages from an immune inhibitory M2-like phenotype toward an immune stimulatory M1-like phenotype and in facilitating CD4(+) and CD8(+) T-cell tumor infiltration . Based on this mechanism , scheduling lower-dose anti-VEGFR2 therapy with T-cell activation induced by a whole cancer cell vaccine therapy enhanced anticancer efficacy in a CD8(+) T-cell-dependent manner in both immune-tolerant and immunogenic murine breast cancer models . These findings indicate that vascular-normalizing lower doses of anti-VEGFR2 antibody can reprogram the tumor microenvironment away from immunosuppression toward potentiation of cancer vaccine therapies . Given that the combinations of high doses of bevacizumab with chemotherapy have not improved overall survival of breast cancer patients , our study suggests a strategy to use antiangiogenic agents in breast cancer more effectively with active immunotherapy and potentially other anticancer therapies . OUTPUT: inducing angiogenesis;avoiding immune destruction INPUT: Regulation of adaptive immunity by innate immune cells is widely accepted . Conversely , adaptive immune cells can also regulate cells of the innate immune system . Here , we report for the first time the essential role of B cells in regulating macrophage ( Mφ ) phenotype . In vitro B cell/Mφ co-culture experiments together with experiments in transgenic mice models for B-cell deficiency or overexpression showed B1 cells to polarize Mφ to a distinct phenotype . This was characterized by downregulated TNF-α , IL-1β and CCL3 , but upregulated IL-10 upon LPS stimulation ; constitutive expression of M2 Mφ markers ( e.g . Ym1 , Fizz1 ) and overexpression of TRIF-dependent cytokines ( IFN-β , CCL5 ) . Mechanistically , this phenotype was linked to a defective NF-κB activation , but a functional TRIF/STAT1 pathway . B1-cell-derived IL-10 was found to be instrumental in the polarization of these Mφ . Finally , in vivo relevance of B1-cell-induced Mφ polarization was confirmed using the B16 melanoma tumor model where adoptive transfer of B1 cells induced an M2 polarization of tumor-associated Mφ . Collectively , our results define a new mechanism of Mφ polarization wherein B1 cells play a key role in driving Mφ to a unique , but M2-biased phenotype . Future studies along these lines may lead to targeting of B1 cells to regulate Mφ response in inflammation and cancer . OUTPUT: tumor promoting inflammation INPUT: beta-Defensins are small antimicrobial peptides of the innate immune system produced in response to microbial infection of mucosal tissue and skin . We demonstrate that murine beta-defensin 2 ( mDF2beta ) acts directly on immature dendritic cells as an endogenous ligand for Toll-like receptor 4 ( TLR-4 ) , inducing up-regulation of costimulatory molecules and dendritic cell maturation . These events , in turn , trigger robust , type 1 polarized adaptive immune responses in vivo , suggesting that mDF2beta may play an important role in immunosurveillance against pathogens and , possibly , self antigens or tumor antigens . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot20

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: PURPOSE Current antibody-based immunotherapeutic approaches under evaluation for breast carcinoma are limited in target scope . For example , administration of the human epidermal growth factor receptor ( EGFR ) antibody , alone or in combination with a chemotherapeutic drug , is thought to primarily inhibit tumor cell proliferation . The aim of this study was to assess the effects of a combined blockade designed to inhibit tumor growth by inhibition of proliferation rate and the proinflammatory effects of interleukin ( IL ) 8 . EXPERIMENTAL DESIGN A human breast carcinoma cell line that produces high levels of IL-8 was injected s.c. into severe combined immunodeficient mice . IL-8 has been reported to augment the progression of some human tumors ; thus , we used a human IL-8 antibody , ABXIL8 , in combination with anti-EGFR , ABXEGFR , to inhibit the metastasis of MDA231 tumors . RESULTS Whereas anti-IL-8 alone had no appreciable antimetastatic effect , the combination of ABXIL8 significantly enhanced the antitumor effects of ABXEGFR , resulting in greater survival of SCID tumor-bearing mice . This effect on survival was correlated with decreased metastatic spread and decreased tumor size in mice receiving both antibodies . Intriguingly , in vitro studies indicate that this antibody combination markedly inhibited matrix metalloproteinase activity associated with MDA-231 cells to a greater degree than either antibody alone . CONCLUSION Combined administration of these two human antibodies using growth factor blockade in conjunction with chemokine blockade may thus provide a more effective approach for treatment of metastatic human breast carcinoma . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND Adult human mesenchymal stem cells ( hMSC ) have been shown to home to sites of carcinoma and affect biological processes , including tumour growth and metastasis . Previous findings have been conflicting and a clear understanding of the effects of hMSCs on cancer remains to be established . Therefore , we set out to investigate the impact of hMSCs on the oestrogen receptor positive , hormone-dependent breast carcinoma cell line MCF-7 . RESULTS In this study , we show the effects of hMSCs on cancer cells are mediated through a secreted factor(s) which are enhanced by cancer cell-hMSC contact/communication . In addition to enhanced proliferation when in co-culture with hMSCs , MCF-7 cells were found to have increased migration potential in vitro . Inhibition of ER signalling by the pure anti-oestrogen ICI 182,780 decreased the effect of hMSCs on MCF-7 cell proliferation and migration supporting a role for ER signalling in the hMSC/MCF-7 cell interaction . Additionally , hMSCs have been shown to secrete a wide variety of growth factors and chemokines including stromal cell-derived factor-1 ( SDF-1 ) . This coupled with the knowledge that SDF-1 is an ER-mediated gene linked with hormone-independence and metastasis led to the investigation of the SDF-1/CXCR4 signalling axis in hMSC-MCF-7 cell interaction . Experiments revealed an increase in SDF-1 gene expression both in vivo and in vitro when MCF-7 cells were cultured with hMSCs . SDF-1 treatment of MCF-7 cells alone increased proliferation to just below that seen with hMSC co-culture . Additionally , blocking SDF-1 signalling using a CXCR4-specific inhibitor decreased hMSC induced proliferation and migration of MCF-7 . However , the combined treatment of ICI and AMD3100 reduced MCF-7 cell proliferation and migration below control levels , indicating targeting both the ER and CXCR4 pathways is effective in decreasing the hMSCs induction of MCF-7 cell proliferation and migration . CONCLUSIONS The sum of these data reveals the relationship between tumour microenvironment and tumour growth and progression . Better understanding of the mechanisms involved in this tumour stroma cell interaction may provide novel targets for the development of treatment strategies for oestrogen receptor positive , hormone-independent , and endocrine-resistant breast carcinoma . OUTPUT: activating invasion and metastasis;sustaining proliferative signaling INPUT: Energy deregulation and abnormalities of tumor cell metabolism are critical issues in understanding cancer . Hereditary leiomyomatosis renal cell carcinoma ( HLRCC ) is an aggressive form of RCC characterized by germline mutation of the Krebs cycle enzyme fumarate hydratase ( FH ) , and one known to be highly metastatic and unusually lethal . There is considerable utility in establishing preclinical cell and xenograft models for study of disorders of energy metabolism , as well as in development of new therapeutic approaches targeting of tricarboxylic acid ( TCA ) cycle enzyme-deficient human cancers . Here we describe a new immortalized cell line , UOK 262 , derived from a patient having aggressive HLRCC-associated recurring kidney cancer . We investigated gene expression , chromosome profiles , efflux bioenergetic analysis , mitochondrial ultrastructure , FH catabolic activity , invasiveness , and optimal glucose requirements for in vitro growth . UOK 262 cells have an isochromosome 1q recurring chromosome abnormality , i(1)(q10) , and exhibit compromised oxidative phosphorylation and in vitro dependence on anaerobic glycolysis consistent with the clinical manifestation of HLRCC . The cells also display glucose-dependent growth , an elevated rate of lactate efflux , and overexpression of the glucose transporter GLUT1 and of lactate dehydrogenase A ( LDHA ) . Mutant FH protein was present primarily in edematous mitochondria , but with catalytic activity nearly undetectable . UOK 262 xenografts retain the characteristics of HLRCC histopathology . Our findings indicate that the severe compromise of oxidative phosphorylation and rapid glycolytic flux in UOK 262 are an essential feature of this TCA cycle enzyme-deficient form of kidney cancer . This tumor model is the embodiment of the Warburg effect . UOK 262 provides a unique in vitro and in vivo preclinical model for studying the bioenergetics of the Warburg effect in human cancer . OUTPUT: cellular energetics INPUT: OBJECTIVE Overexpression of epidermal growth factor receptor ( EGFR ) in glioblastoma multiforme ( GBM ) secondary to EGFR gene amplification is associated with a more aggressive tumor phenotype and a worse clinical outcome . The purpose of this study was to analyze whether blocking this receptor with the anti-EGFR chimeric monoclonal antibody C225 would decrease proliferation and increase apoptosis in GBM cells . METHODS EGFR expression and amplification were analyzed for seven human GBM cell lines . These lines were then exposed to different concentrations of C225 for 48 hours , 72 hours , and 7 days , after which time cytotoxicity , apoptosis , and vascular endothelial growth factor expression were assessed in vitro . Two EGFR-amplified human GBM were implanted in the flanks of nude mice , and the animals received C225 twice per week intraperitoneally for 5 weeks . Tumor volumes and survival times were compared with those of sham-treated mice . RESULTS EGFR gene amplification was demonstrated in three of the primary GBM lines . C225 treatment produced significant cytotoxicity in all three EGFR-amplified GBM lines , but not in unamplified lines . Flow cytometry demonstrated increased apoptosis in C225-treated , EGFR-amplified GBM lines , but not in unamplified lines . There was a decrease in vascular endothelial growth factor expression in all GBM lines with exposure to C225 . Tumor-bearing mice treated with C225 experienced significant inhibition of tumor growth as well as a 200% increase in median survival . CONCLUSION Blocking EGFR in GBM cells that overexpress this receptor significantly changes tumor cell biology by promoting apoptosis while decreasing proliferation and vascular endothelial growth factor expression . This approach holds great promise for the treatment of patients with GBMs . OUTPUT: sustaining proliferative signaling;resisting cell death;inducing angiogenesis INPUT: γ-Tocotrienol and sesamin are phytochemicals that display potent anticancer activity . Since sesamin inhibits the metabolic degradation of tocotrienols , studies were conducted to determine if combined treatment with sesamin potentiates the antiproliferative effects of γ-tocotrienol on neoplastic mouse ( +SA ) and human ( MCF-7 and MDA-MB-231 ) mammary cancer cells . Results showed that treatment with γ-tocotrienol or sesamin alone induced a significant dose-responsive growth inhibition , whereas combination treatment with these agents synergistically inhibited the growth of +SA , MCF-7 and MDA-MB-231 mammary cancer cells , while similar treatment doses were found to have little or no effect on normal ( mouse CL-S1 and human MCF-10A ) mammary epithelial cell growth or viability . However , sesamin synergistic enhancement of γ-tocotrienol-induced anticancer effects was not found to be mediated from a reduction in γ-tocotrienol metabolism . Rather , combined treatment with subeffective doses of γ-tocotrienol and sesamin was found to induce G1 cell cycle arrest , and a corresponding decrease in cyclin D1 , CDK2 , CDK4 , CDK6 , phospho-Rb , and E2F1 levels , and increase in p27 and p16 levels . Additional studies showed that the antiproliferative effect of combination treatment did not initiate apoptosis or result in a decrease in mammary cancer cell viability . Taken together , these findings indicate that the synergistic antiproliferative action of combined γ-tocotrienol and sesamin treatment in mouse and human mammary cancer cells is cytostatic , not cytotoxic , and results from G1 cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: PURPOSE To investigate treatment of human pancreatic cancer cell lines and xenografts with combinations of Erbitux ( IMC-C225 ) anti-epidermal growth factor receptor ( EGFR ) antibody , gemcitabine , and radiation . METHODS AND MATERIALS BxPC-3 and MiaPaCa-2 human pancreatic carcinoma cells were treated in vitro for 24 h with IMC-C225 ( 5 microg/mL ) , then exposed to epidermal growth factor ( EGF ) ( 10 mM ) for 5 min . Immunoblots were screened for EGFR expression and the ability of IMC-C225 to block EGF-induced tyrosine phosphorylation of EGFR . Cells were treated with IMC-C225 ( 5 microg/mL ) on Day 0 , the IC(50) dose of gemcitabine on Day 1 for 24 h , followed by 3 Gy 60Co irradiation on Day 2 , or the combination of each agent . For cell proliferation , cells were counted on Day 4 , and for apoptosis , cells were stained with annexin V-FITC and propidium iodide , then analyzed by FACS . Cells were treated with the same single or multiple treatments and analyzed in a clonogenic cell survival assay . The effect of IMC-C225 , gemcitabine , and radiation on the growth of BxPC-3 and MiaPaCa-2 tumor xenografts was determined . Athymic nude mice bearing established s.c. tumor xenografts of 6-8 mm diameter received 6 weeks of treatment with IMC-C225 ( 1 mg every 3 days x 6 ) alone or in combination with gemcitabine ( 120 mg/kg i.v. every 6 days x 6 ) , and 6 weekly fractions of 3 Gy radiation on the days after gemcitabine administration . Tumor growth was measured with Vernier calipers . RESULTS BxPC-3 and MiaPaCa-2 cell lines expressed low levels of EGFR . IMC-C225 inhibited EGF-induced tyrosine phosphorylation of the EGF receptor on both cell lines . Treatment of cells with a combination of IMC-C225 + gemcitabine + radiation produced the highest induction of apoptosis and inhibition of proliferation in vitro . Combination treatment with IMC-C225 , gemcitabine , and radiation produced 100% complete regression of MiaPaCa-2 tumors for more than 250 days , and the greatest growth inhibition of BxPC-3 tumors compared to any single or dual treatments . CONCLUSIONS The IMC-C225 therapy in combination with gemcitabine chemotherapy and radiation therapy demonstrated statistically significantly greater efficacy over the single and double combination therapies . This form of multimodality treatment shows potential clinical application in the treatment of pancreatic cancer in humans . OUTPUT:

sustaining proliferative signaling;resisting cell death

HoC_dynamic_5_shot21

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: UVB from solar radiation is both an initiating and promoting agent for skin cancer . We have found that primary human keratinocytes undergo an apoptotic response to UVB . To determine whether these responses are altered during the course of immortalization , we examined markers of apoptosis in primary human foreskin keratinocytes ( HFK ) transduced with either a retroviral vector expressing the E6 and E7 genes of HPV-16 or with empty vector alone ( LXSN-HFK ) . Whereas LXSN-HFK as well as early passage keratinocytes expressing HPV-16 E6 and E7 ( p7 E6/7-HFK ) were both moderately responsive to UVB irradiation , late passage-immortalized keratinocytes ( p27 E6/7-HFK ) were exquisitely sensitive to UVB-induced apoptosis . After exposure to UVB , enhanced annexin V-positivity and internucleosomal DNA fragmentation were observed in p27 E6/7-HFK compared with either LXSN- or p7 E6/7-HFK . Caspase-3 fluorometric activity assays as well as immunoblot analysis with antibodies to caspase-3 and poly(ADP-ribose) polymerase revealed elevated caspase-3 activity and processing at lower UVB doses in p27 E6/7-HFK compared with LXSN- or p7 E6/7-HFK . In addition , the caspase inhibitor DEVD-CHO reduced the apoptotic response and increased survival of all three HFK types . Immunoblot analysis revealed that caspase-8 was activated in all three cell types , but caspase-9 was only activated in p27 E6/7-HFK . Cell cycle analysis further showed that only p27 E6/7-HFK exhibit G(2)/M accumulation that is enhanced by UVB treatment . This accumulation was associated with a rapid down-regulation of Bcl-2 in these cells . The immortalization process subsequent to the expression of HPV E6 and E7 may therefore determine UVB sensitivity by switching the mode of apoptosis from a caspase-8 to a Bcl-2-caspase-9-mediated pathway of apoptosis . OUTPUT: resisting cell death;enabling replicative immortality INPUT: Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers . A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation . We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products . This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine ( NMBzA ) , and the response that increased levels of dietary vitamin E has on this induced carcinogenesis . Female C57BL/6 mice received NMBzA or vehicle ( corn oil ) i.p. weekly for 3 weeks . Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet . As an assessment of free radical activity , exhaled ethane was measured prior to killing the animals at 26 weeks . Esophagi from the various mice groups were assessed for size and frequency of tumors . Livers homogenates were analyzed for vitamins A and E , lipid fluorescence , conjugated dienes and malondialdehyde . Hepatic levels of vitamin A and E were decreased ( P &lt ; 0.05 ) and indices of lipid peroxidation were greater ( P &lt ; 0.05 ) in NMBzA-treated mice relative to controls . Lipid peroxidation and serum transaminases ( ALT and AST ) were greatest in mice given NMBzA and infected with the retroviruses . Incidence of esophageal tumors were also greatest in the NMBzA-treated , immunocompromised animals . Mice fed vitamin E-supplemented diets showed increased ( P &lt ; 0.05 ) hepatic concentrations of vitamin E and vitamin A , decreased activities of serum transaminases , decreased indices of lipid peroxidation , and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice . These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals . OUTPUT: avoiding immune destruction INPUT: One of the immunosuppressive effects of both ultraviolet ( UV ) light and chemical carcinogens is to deplete Langerhans cells ( LC ) from the epidermis , suggesting that these cells play an important role in inducing immune responses to developing tumors during the early phases of carcinogenesis . Retinoids such as all-trans-retinoic acid ( RA ) are natural or synthetic derivatives of vitamin A ; RA binds to nuclear receptors in the skin , effecting transcription of a wide range of genes . Topical application of RA prevents the tumor promotor 12-O-tetradecanoylphorbol-13-acetate ( TPA ) from depleting the density of LC in murine epidermis . In contrast , topical RA did not itself alter the normal LC density . RA also inhibited the development of TPA-induced immunosuppression to a locally applied contact sensitizer . Topical RA also prevented UV light from reducing the density of both LC and Thy-1+ dendritic epidermal cells ( Thy-1+ dEC ) . However , the RA treatment did not prevent local immunosuppression to the contact sensitizer from developing in response to UV irradiation . The reasons for this are unclear , however , it is possible that RA does not inhibit some other immunosuppressive effect of UV light . Temarotene , a recently developed synthetic retinoid also inhibited UV light from reducing the LC and Thy-1+ dEC density from murine epidermis . Thus part of the anti-carcinogenic activity of retinoids may be due to their ability to protect LC during the early stages of carcinogenesis . OUTPUT: avoiding immune destruction INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Nitric oxide ( NO)-releasing non-steroidal anti-inflammatory drugs ( NO-NSAIDs ) which have been synthesized to reduce gastro-intestinal and cardiovascular toxicities of NSAIDs , possess anti-proliferative , pro-apoptotic and anti-cancer activities . Here , we show that NO-sulindac inhibited UVB-induced skin tumorigenesis in SKH-1 hairless mice . Topical application of NO-sulindac reduced tumor incidence , number ( p<0.05 ) and volume ( p<0.005 ) as compared to UVB ( alone)-irradiated vehicle-treated mice . An increase in TUNEL-positive cells in skin lesions was accompanied by the enhanced Bax:Bcl-2 ratio . The expression of pro-apoptotic Bax was increased whereas anti-apoptotic Bcl-2 reduced . However , proliferation was identified as the major target of NO-sulindac in this study . A reduced expression of PCNA and cyclin D1 associated with the dampening of cell cycle progression was observed . The mechanism of this inhibition was related to the reduction in UVB-induced Notch signaling pathway . UVB-induced inflammatory responses were diminished by NO-sulindac as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases Erk1/2 , p38 and JNK1/2 . In this regard , NO-sulindac also inhibited NFκB by enhancing IκBα as evidenced by the reduced expression of iNOS and COX-2 , the direct NFκB transcription target proteins . NO-sulindac significantly diminished the progression of benign lesions to invasive carcinomas by suppressing the tumor aggressiveness and retarding epithelial-mesenchymal transition . A marked decrease in the expression of mesenchymal markers such as Fibronectin , N-cadherin , SNAI , Slug and Twist and an increase in epithelial cell polarity marker E-cadherin were noted in NO-sulindac-treated tumors . Our data suggest that NO-sulindac is a potent inhibitor of UVB-induced skin carcinogenesis and acts by targeting proliferation-regulatory pathways . OUTPUT: resisting cell death;sustaining proliferative signaling;tumor promoting inflammation;activating invasion and metastasis INPUT: The ultraviolet ( UV ) radiation present in sunlight is immune-suppressive . Recently we showed that solar-simulated UV radiation ( UVA + UVB ; 295-400 nm ) , applied after immunization , suppressed immunological memory and the elicitation of delayed-type hypersensitivity to the common opportunistic pathogen , Candida albicans . Further , we found that wavelengths in the UVA region of the solar spectrum ( 320-400 nm ) , devoid of UVB , were equally effective in activating immune suppression as UVA + UVB radiation . Here we report on the mechanisms involved . No immune suppression was found in UV-irradiated mice injected with monoclonal anti-interleukin ( IL)-10 antibody , or mice exposed to solar-simulated UV radiation and injected with recombinant IL-12 . Antigen-specific suppressor T cells were found in the spleens of mice exposed to UVA + UVB radiation . Applying liposomes containing bacteriophage T4N5 to the skin of mice exposed to solar-simulated UVA + UVB radiation or mice exposed to UVA radiation blocked immune suppression , demonstrating an essential role for UV-induced DNA damage in the suppression of established immune reactions . These findings indicate that UV radiation activates similar immunological pathways to suppress the induction , or the elicitation , of the immune response . OUTPUT:

avoiding immune destruction;genomic instability and mutation

HoC_dynamic_5_shot22

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Androgens and the androgen receptor ( AR ) play important roles in the development of male urogenital organs . We previously found that mice with total AR knockout ( ARKO ) and epithelial ARKO failed to develop normal prostate with loss of differentiation . We have recently knocked out AR gene in smooth muscle cells and found the reduced luminal infolding and IGF-1 production in the mouse prostate . However , AR roles of stromal fibroblasts in prostate development remain unclear . METHODS To further probe the stromal fibroblast AR roles in prostate development , we generated tissue-selective knockout mice with the AR gene deleted in stromal fibroblasts ( FSP-ARKO ) . We also used primary culture stromal cells to confirm the in vivo data and investigate mechanisms related to prostate development . RESULTS The results showed cellular alterations in the FSP-ARKO mouse prostate with decreased epithelial proliferation , increased apoptosis , and decreased collagen composition . Further mechanistic studies demonstrated that FSP-ARKO mice have defects in the expression of prostate stromal growth factors . To further confirm these in vivo findings , we prepared primary cultured mouse prostate stromal cells and found knocking down the stromal AR could result in growth retardation of prostate stromal cells and co-cultured prostate epithelial cells , as well as decrease of some stromal growth factors . CONCLUSIONS Our FSP-ARKO mice not only provide the first in vivo evidence in Cre-loxP knockout system for the requirement of stromal fibroblast AR to maintain the normal development of the prostate , but may also suggest the selective knockdown of stromal AR might become a potential therapeutic approach to battle prostate hyperplasia and cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Cellular senescence is considered as a tumor suppressive mechanism . Recent evidence indicates however that senescent cells secrete various growth factors and cytokines , some of which may paradoxically promote cancer progression . This phenomenon termed senescence-associated secretory phenotype ( SASP ) must be inhibited in order for anti-proliferative agents to be effective . The present study was designed to determine whether the β-catenin destruction complex ( BCDC ) , known to integrate the action of various growth factors and cytokines , would represent a suitable target to inhibit the activity of SASP components . For this , we carried out experiments to determine the effect of drug-induced senescence on secretion of SASP , β-catenin transactivation , and the relationship between these processes . Moreover , genetic and pharmacological approaches were used to define the implication of BCDC in mediating the effects of SASP components on cell migration and resistance to drugs . The findings indicate that drug-induced senescence was associated with expression of various Wnt ligands in addition to previously known SASP components . Beta catenin transactivation and expression of genes implicated in epithelial-mesenchymal transition ( EMT ) also increased in response to drug-induced SASP . These effects were prevented by Pyrvinium , a recently described activator of BCDC . Pyrvinium also suppressed the effects of SASP on cell migration and resistance to doxorubicin . Together , these findings provide insights on the potential role of BCDC in mediating the effects of drug-induced SASP on cancer cell invasion and resistance to therapy , and suggest that targeting this pathway may represent an effective approach to enhance the activity of current and prospective anti-cancer therapeutics . OUTPUT: enabling replicative immortality;activating invasion and metastasis INPUT: OBJECTIVES Epiphyseal cartilage is a barrier to osteosarcoma invasion , however the mechanisms behind this resistance remain unclear . The aim of this study was to examine the chronological and spatial patterns of osteosarcoma growth and invasion of local tissue structures including epiphyseal cartilage . METHODS We used an in vivomouse model of osteosarcoma to histologically examine tumors at different stages of disease progression . We compared the pattern of osteosarcoma penetration of epiphyseal cartilage with the expression pattern of two potent mediators of angiogenesis ; proangiogenic vascular endothelial growth factor ( VEGF ) and antiangiogenic pigment epithelium-derived factor ( PEDF ) . RESULTS Epiphyseal cartilage remained intact across its entire length in all sections examined , despite increasing tumor size as well as intra- and extraosseous destruction . In the most advanced cases , only the proangiogenic lowermost layers of the hypertrophic zone of the growth plate were eroded . This corresponded with the growth plate layers which highly expressed the angiogenic factor VEGF . In contrast , the resting , proliferative and upper hypertrophic layers were resistant to osteosarcoma invasion in all cases . This corresponded to the layers with the highest expression of the potent antiangiogenic factor PEDF . CONCLUSION Epiphyseal cartilage is resistant to local invasion by osteosarcoma . The balance of angiogenesis , influenced by pro- and antiangiogenic factors , is likely to play an important role in this resistance . OUTPUT: inducing angiogenesis;activating invasion and metastasis INPUT: Annexin II is secreted into the extracellular environment , where , via interactions with specific proteases and extracellular matrix proteins , it participates in plasminogen activation , cell adhesion , and tumor metastasis and invasion . However , mechanisms regulating annexin II transport across the cellular membrane are unknown . In this study , we used coimmunoprecipitation to show that Annexin-II was bound to insulin and insulin-like growth factor-1 ( IGF-1 ) receptors in PC12 cells and NIH-3T3 cells overexpressing insulin ( NIH-3T3(IR) ) or IGF-1 receptor ( NIH-3T3(IGF-1R) ) . Stimulation of insulin and IGF-1 receptors by insulin caused a temporary dissociation of annexin II from these receptors , which was accompanied by an increased amount of extracellular annexin II detected in the media of PC12 , NIH-3T3(IR) , and NIH-3T3(IGF-1R) cells but not in that of untransfected NIH-3T3 cells . Activation of a different growth factor receptor , the platelet-derived growth factor receptor , did not produce such results . Tyrphostin AG1024 , a tyrosine kinase inhibitor of insulin and IGF-1 receptor , was shown to inhibit annexin II secretion along with reduced receptor phosphorylation . Inhibitors of a few downstream signaling enzymes including phosphatidylinositol 3-kinase , pp60c-Src , and protein kinase C had no effect on insulin-induced annexin II secretion , suggesting a possible direct link between receptor activation and annexin II secretion . Immunocytochemistry revealed that insulin also induced transport of the membrane-bound form of annexin II to the outside layer of the cell membrane and appeared to promote cell aggregation . These results suggest that the insulin receptor and its signaling pathways may participate in molecular mechanisms mediating annexin II secretion . OUTPUT: sustaining proliferative signaling INPUT: Peptide growth factors have been implicated in progression of prostate cancer ( PCa ) to the androgen-independent state ; however , much of the evidence linking diffusible mitogens and survival factors to this process remains circumstantial . Heparin-binding epidermal growth factor-like growth factor ( HB-EGF ) , a prostate stroma-derived factor , promotes survival , proliferation , and neuroendocrine differentiation of androgen-dependent LNCaP PCa cells in vitro . To test whether sustained exposure to HB-EGF can confer an androgen-independent phenotype , we generated stable populations of LNCaP cells that express constitutively a secreted form of HB-EGF ( LNCaP/sHB ) . LNCaP/sHB cells proliferated more rapidly under androgen-depleted conditions in vitro and formed larger tumors with higher frequency in intact and castrated severe combined immunodeficient mice , in comparison to control cells . LNCaP/sHB tumors also expressed higher levels of the neuroendocrine marker , neuron-specific enolase , compared with control tumors . In castrates , increased neuron-specific enolase expression in LNCaP/sHB tumors was associated with reduced androgen receptor ( AR ) levels . In vitro , AR protein levels were reduced in LNCaP/sHB cells , and in transient transfection assays using an androgen-responsive promoter ( mouse mammary tumor virus-long terminal repeat ) , LNCaP/sHB cells showed reduced sensitivity to dihydrotestosterone compared with controls . This is the first demonstration that continuous exposure of AR-positive PCa cells to a single growth factor can promote an androgen-independent phenotype in vivo . These findings also emphasize the potential role of pathways other than the AR axis in acquisition of androgen independence . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot23

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: BACKGROUND The pTalpha/preTCR regulates the beta-selection , a crucial T-cell developmental checkpoint , providing a most potent survival advantage to thymocytes mediated by the src-kinase p56(Lck) . METHODS To define the relevance of pTalpha in human T-cell lymphoblastic leukemia ( T-ALL ) , we analyzed in T-ALL cell lines ( n=14 ) pTalpha and p56(Lck) mRNA and protein expression as also the tyrosine-phosphorylation . The p56(Lck) specific src-protein-tyrosine kinase inhibitor ( PTK-I ) PP1 was used in growth inhibition assays . IC(50) value determination , cell cycle- and apoptosis analyses were performed in T-ALL- , non-T-ALL- and murine transgenic cell lines . RESULTS pTalpha expression patterns were markedly different in T-ALL cell lines as compared to those reported for normal lymphoid counterparts . PP1 induced in 6/11 T-ALL cell lines a survival disadvantage resulting from a cell cycle arrest in the G(1/0) phase in thymic lymphoblastic cells and apoptosis induction in the immature cell line HSB-2 , respectively . PP1 sensitive cell lines expressed the target protein p56(Lck) and showed a corresponding P-Tyr signal . CONCLUSION Sensitivity of thymic T-ALLs to PP1 clearly underlines the impact of pTalpha mediated proliferation in this leukemic sub-type . In addition , p56(Lck) represents also independently of pTalpha a promising therapeutical target for the src-kinase inhibitors in neoplastic lymphoid diseases . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Abstract Purpose : Cell death is one of the most important endpoints of radiosensitivity . The tumor suppressor p53 participates not only in regulation of apoptosis , but also in autophagy mechanism . In this study , H1299-P53 ( with wild-type p53 ) and H1299-175H ( with mutant 175H ) were used , and the effects of p53 on radiosensitivity were analyzed . Methods : Cell models with different p53 status were established by gene engineering , and cell viability was examined by colony formation assay , and cell counting kit-8 ( CCK-8 ) , 3-Methyladenine , and Z-VAD were used to block autophagy and apoptosis , respectively . Western blot was used to detect protein expression ; monodansylcadaverine ( MDC ) staining was used to analyze autophagy rate ; DAPI/Propidium Iodide ( PI ) staining and flow cytometry were used to assess apoptosis and necrosis . Results : In parental H1299 , H1299-P53 , and H1299-175H cells , radiosensitivity exhibited different by colony formation and CCK-8 assay ( D0 : 1.764 Gy , 1.407 Gy and 1.695 Gy ; Dq : 2.977 Gy , 1.199 Gy and 2.312 Gy in turn ) . The radiosensitization of p53 was associated with the increase of MDM2 and P21 expression . The ionizing radiation ( IR)-induced apoptosis was significant in H1299-P53 compared with in H1299 and H199-175H ( p<0.05 ) by flow cytometry , and the expression of cleaved-caspase3 was increased in H1299-P53 cells . While the IR-induced autophagy was significant in H1299 cells ( p<0.01 ) and decreased in H1299-P53 and H1299-175H cells ( p<0.01 ) by MDC staining , the expression of MAPLC3II and Beclin-1 increased in H1299 , but not in H1299-p53 and H199-175H cells . The IR-induced cell survival was significantly increased by Z-VAD-FMK and decreased by 3MA in H1299-P53 cells ; IR- induced autophagy was significantly increased by Z-VAD-FMK in H1299-P53 cells ( p<0.01 ) , but not changed in H1299 cells . Conclusion : p53 could regulate radiosensitivity by inhibiting autophagy and activating apoptosis ; autophagy provides a prosurvival mechanism , and p53 potently abrogated the IR-induced autophagy , while mutant 175H shown no effect on radiosensitivity , suggesting that individual treatment strategies should be based on p53 status in patients . OUTPUT: resisting cell death INPUT: The influence of radiation-induced apoptosis on radiosensitivity was studied in a set of closely related human lymphoblastoid cell lines differing in TP53 status . The clonogenic survival of irradiated TK6 cells ( expressing wild-type TP53 ) , WTK1 cells ( overexpressing mutant TP53 ) , and TK6E6 cells ( negative for TP53 owing to transfection with HPV16 E6 ) was assessed in relation to the induction of apoptosis and its suppression by caspase inhibition or treatment with PMA as well as after treatment with caffeine . Measurements using the alkaline comet assay and pulsed-field electrophoresis of the induction and repair of DNA strand breaks showed similar kinetics of the processing of early DNA damage in these cell lines . The cytochalasin B micronucleus assay revealed identical levels of residual damage in the first postirradiation mitosis of these cells . Abrogation of TP53-dependent apoptosis in TK6E6 cells resulted in a distinct increase in radioresistance . Further suppression of apoptosis as observed in WTK1 cells overexpressing mutant TP53 apparently was not responsible for the high radioresistance of WTK1 cells , since other means of highly efficient suppression of apoptosis ( caspase inhibition or PMA treatment ) increased the clonogenic survival of irradiated TK6 cells only to levels similar to those of TK6E6 cells with abrogated TP53-dependent apoptosis . Considering the similar levels of residual chromosomal damage in TK6E6 cells and WTK1 cells , a hitherto unknown mechanism of tolerance needs to be inferred for these TP53 mutant cells . This residual damage tolerance , however , appears to require an intact G2/M-phase checkpoint function since the relative radioresistance of the WTK1 cells was completely lost upon caffeine treatment , which also resulted in a failure of the TK6 and TK6E6 cells to execute apoptosis . In this situation , the cellular response seems to be dominated entirely by TP53-independent mitotic failure . OUTPUT:

genomic instability and mutation;resisting cell death

HoC_dynamic_5_shot24

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: BACKGROUND Death receptors ( DR ) of the TNF family function as anti-tumor immune effector molecules . Tumor cells , however , often exhibit DR-signaling resistance . Previous studies indicate that radiation can modify gene expression within tumor cells and increase tumor cell sensitivity to immune attack . The aim of this study is to investigate the synergistic effect of sub-lethal doses of ionizing radiation in sensitizing colorectal carcinoma cells to death receptor-mediated apoptosis . METHODOLOGY/PRINCIPAL FINDINGS The ability of radiation to modulate the expression of multiple death receptors ( Fas/CD95 , TRAILR1/DR4 , TRAILR2/DR5 , TNF-R1 and LTβR ) was examined in colorectal tumor cells . The functional significance of sub-lethal doses of radiation in enhancing tumor cell susceptibility to DR-induced apoptosis was determined by in vitro functional sensitivity assays . The longevity of these changes and the underlying molecular mechanism of irradiation in sensitizing diverse colorectal carcinoma cells to death receptor-mediated apoptosis were also examined . We found that radiation increased surface expression of Fas , DR4 and DR5 but not LTβR or TNF-R1 in these cells . Increased expression of DRs was observed 2 days post-irradiation and remained elevated 7-days post irradiation . Sub-lethal tumor cell irradiation alone exhibited minimal cell death , but effectively sensitized three of three colorectal carcinoma cells to both TRAIL and Fas-induced apoptosis , but not LTβR-induced death . Furthermore , radiation-enhanced Fas and TRAIL-induced cell death lasted as long as 5-days post-irradiation . Specific analysis of intracellular sensitizers to apoptosis indicated that while radiation did reduce Bcl-X(L) and c-FLIP protein expression , this reduction did not correlate with the radiation-enhanced sensitivity to Fas and/or TRAIL mediated apoptosis among the three cell types . CONCLUSIONS/SIGNIFICANCE Irradiation of tumor cells can overcome Fas and TRAIL resistance that is long lasting . Overall , results of these investigations suggest that non-lethal doses of radiation can be used to make human tumors more amenable to attack by anti-tumor effector molecules and cells . OUTPUT: resisting cell death INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: The role of regulatory T cells ( T(regs) ) in human colon cancer ( CC ) remains controversial : high densities of tumor-infiltrating T(regs) can correlate with better or worse clinical outcomes depending on the study . In mouse models of cancer , T(regs) have been reported to suppress inflammation and protect the host , suppress T cells and protect the tumor , or even have direct cancer-promoting attributes . These different effects may result from the presence of different T(reg) subsets . We report the preferential expansion of a T(reg) subset in human CC with potent T cell-suppressive , but compromised anti-inflammatory , properties ; these cells are distinguished from T(regs) present in healthy donors by their coexpression of Foxp3 and RORγt . T(regs) with similar attributes were found to be expanded in mouse models of hereditary polyposis . Indeed , ablation of the RORγt gene in Foxp3(+) cells in polyp-prone mice stabilized T(reg) anti-inflammatory functions , suppressed inflammation , improved polyp-specific immune surveillance , and severely attenuated polyposis . Ablation of interleukin-6 ( IL-6 ) , IL-23 , IL-17 , or tumor necrosis factor-α in polyp-prone mice reduced polyp number but not to the same extent as loss of RORγt . Surprisingly , loss of IL-17A had a dual effect : IL-17A-deficient mice had fewer polyps but continued to have RORγt(+) T(regs) and developed invasive cancer . Thus , we conclude that RORγt has a central role in determining the balance between protective and pathogenic T(regs) in CC and that T(reg) subtype regulates inflammation , potency of immune surveillance , and severity of disease outcome . OUTPUT: tumor promoting inflammation INPUT: Cytokines are known to play an important role in host defense by regulating the function , growth , and differentiation of the cells of the immune system . We hypothesize that , in the tumor microenvironment , tumor cells and resident tissue cells ( e.g. , fibroblasts ) also produce cytokines that may regulate the local immune response to tumors . Initially , homogenates of eight head and neck squamous cell carcinomas ( HNSCC ) were assayed for the presence of interleukin-1 ( IL-1 ) , interleukin-4 ( IL-4 ) , interleukin-6 ( IL-6 ) , and granulocyte-macrophage colony-stimulating factor ( GM-CSF ) to establish the presence of these cytokines in the tumors in vivo . We detected IL-1 in all tumor homogenates and IL-4 , IL-6 , and GM-CSF in some homogenates . To assess the ability of HNSCC to produce these cytokines , supernatants of short-term primary cultures of HNSCC were assayed for the same cytokines . No IL-1 was detected , although baseline levels of IL-4 , IL-6 , and GM-CSF were present . However , the stimulation of primary tumor cultures with exogenous IL-1 induced or significantly enhanced production of IL-4 ( p &lt ; 0.01 ) , IL-6 ( p &lt ; 0.001 ) , and GM-CSF ( p &lt ; 0.02 ) . These results support our hypothesis that HNSCC secrete cytokines that may influence the response of local immune cells . Our data also suggest that IL-1 may have a central role in regulating the local immune response through the enhancement or induction of cytokine production by tumor and/or resident tissue cells . OUTPUT: avoiding immune destruction;tumor promoting inflammation INPUT: PURPOSE The roles of terminal sialyl and fucosyl residues in cell surface glycans in the metastatic potential of H7721 cells , a human hepatocarcinoma cell line , were studied . METHODS Neuraminidase and alpha-L-fucosidase were used to remove the sialyl and fucosyl residues , respectively . Cell adhesion to fibronectin ( Fn ) , laminin ( Ln ) , and human umbilical vein epithelial cell ( HUVEC ) , as well as chemotactic cell migration and invasion , were selected as the parameters of metastatic potential ex vivo . RESULTS Sialyl residue is not essential for cell adhesion to Fn , but is important in cell adhesion to Ln and invasion , and is crucial in cell adhesion to HUVEC and migration . In contrast , fucosyl residue contributes more than sialyl residue to cell adhesion to Fn and Ln , but less to adhesion to HUVEC , and is not essential in chemotactic cell migration and invasion . Cell adhesion to HUVEC , migration , and invasion were inhibited by the monoclonal antibody of sialyl Lewis X , but not by the antibody of non-sialyl Lewis X. CONCLUSION Terminal sialyl residues on cell surface glycans are more important than fucosyl residues in mediating cell adhesion to HUVEC and cell migration/invasion , but the reverse is true in cell adhesion to Fn and Ln . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot25

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Genomic instability has long been recognized as the main feature of neoplasia and a factor modulating individual cancer susceptibility . There are attempts to find effective assays of both individual DNA repair capacity and genetic instability , and their relation to the cancer risk . Genetic predisposition plays an important role in the etiology and development of head and neck squamous cell carcinoma ( HNSCC ) . The aim of our study was to search for a correlation between chromosomal instability and DNA repair capacity in HNSCC patients and healthy controls . The chromosomal instability was measured by the number of bleomycin ( BLM)-induced chromosomal aberrations and diepoxybutane ( DEB)-induced sister chromatid exchanges . The DNA repair capacity was assessed using the DEB-induced adaptive response ( AR ) . The HNSCC patients in our study showed a significant increase in chromosomal instability after a preterminal exposure of their lymphocytes to either BLM for the last 5 h or DEB for the last 24 h of incubation . However , the AR was higher in HNSCC patients than in the control group , suggesting an increase in the DNA repair capacity in the cancer patients as compared to the control . There is no correlation between the DNA repair capacity estimated on the basis of preterminal exposures to BLM and DEB and the DNA repair capacity estimated on the basis of the adaptive response to DEB . The preterminal exposure and the adaptive response test may activate different DNA repair mechanisms . OUTPUT: genomic instability and mutation INPUT: INTRODUCTION Various agents used in breast cancer chemotherapy provoke DNA double-strand breaks ( DSBs ) . DSB repair competence determines the sensitivity of cells to these agents whereby aberrations in the repair machinery leads to apoptosis . Proteins required for this pathway can be detected as nuclear foci at sites of DNA damage when the pathway is intact . Here we investigate whether focus formation of repair proteins can predict chemosensitivity of breast cancer . METHODS Core needle biopsy specimens were obtained from sixty cases of primary breast cancer before and 18-24 hours after the first cycle of neoadjuvant epirubicin plus cyclophosphamide ( EC ) treatment . Nuclear focus formation of DNA damage repair proteins was immunohistochemically analyzed and compared with tumor response to chemotherapy . RESULTS EC treatment induced nuclear foci of gammaH2AX , conjugated ubiquitin , and Rad51 in a substantial amount of cases . In contrast , BRCA1 foci were observed before treatment in the majority of the cases and only decreased after EC in thirteen cases . The presence of BRCA1- , gammaH2AX- , or Rad51-foci before treatment or the presence of Rad51-foci after treatment was inversely correlated with tumor response to chemotherapy . DNA damage response ( DDR ) competence was further evaluated by considering all four repair indicators together . A high DDR score significantly correlated with low tumor response to EC and EC + docetaxel whereas other clinicopathological factors analyzed did not . CONCLUSIONS High performing DDR focus formation resulted in tumor resistance to DNA damage-inducing chemotherapy . Our results suggested an importance of evaluation of DDR competence to predict breast cancer chemosensitivity , and merits further studying into its usefulness in exclusion of non-responder patients . OUTPUT: genomic instability and mutation INPUT: DNA repair is an essential cellular process required to maintain genomic stability . Every cell is subjected to thousands of DNA lesions daily under normal physiological conditions . Ionizing radiation ( IR ) is a major DNA damaging agent that can be produced by both natural and man-made sources . A common source of radiation exposure is through its use in medical diagnostics or treatments such as for cancer radiotherapy where relatively high doses are received by patients . To understand the detailed DNA repair gene transcription response to high dose IR , gene expression exon array studies have been performed and the response to radiation in two divergent cell types , lymphoblastoid cell lines and primary fibroblasts , has been examined . These exon arrays detect expression levels across the entire gene , and have the advantage of high sensitivity and the ability to identify alternative transcripts . We found a selection of DNA repair genes , including some not previously reported , that are modulated in response to radiation . Detailed dose and time course kinetics of DNA repair transcription was conducted and results have been validated utilizing PCR methods . Alternative transcription products in response to IR were identified in several DNA repair genes including RRM2B and XPC where alternative initiation sites were found . These investigations have advanced the knowledge about the transcriptional response of DNA repair . OUTPUT: genomic instability and mutation INPUT: When the prostate cancer cells become unresponsive to androgen therapy , resistance to chemotherapy becomes imminent , resulting in high mortality . To combat this situation , lycopodine , a pharmacologically important bioactive component derived from Lycopodium clavatum spores , was tested against hormone sensitive ( LnCaP ) and refractory ( PC3 ) prostate cancer cells in vitro . This study aims to check if lycopodine has demonstrable anti-cancer effects and if it has , to find out the possible mechanism of its action . The MTT assay was performed to evaluate the cytotoxic effect . Depolarization of mitochondrial membrane potential , cell cycle , EGF receptor activity and apoptosis were recorded by FACS ; profiles of different anti- and pro-apoptotic genes and their products were studied by semi-quantitative RT-PCR , indirect-ELISA , western blotting . Drug-DNA interaction was determined by CD spectroscopy . Administration of lycopodine down-regulated the expression of 5-lipoxygenase and the 5-oxo-ETE receptor ( OXE receptor1 ) and EGF receptor , and caused up-regulation of cytochrome c with depolarization of mitochondrial inner membrane potential , without palpable change in p53 activity , resulting in apoptosis , cell arrest at G0/G1 stage and ultimately reduced proliferation of cancer cells ; concomitantly , there was externalization of phosphotidyl serine residues . CD spectroscopic analysis revealed intercalating property of lycopodine with DNA molecule , implicating its ability to block cellular DNA synthesis . The overall results suggest that lycopodine is a promising candidate suitable for therapeutic use as an anti-cancer drug . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Acute endothelial cell apoptosis and microvascular compromise couple gastrointestinal tract irradiation to reproductive death of intestinal crypt stem cell clonogens ( SCCs ) following high-dose radiation . Genetic or pharmacologic inhibition of endothelial apoptosis prevents intestinal damage , but as the radiation dose is escalated , SCCs become directly susceptible to an alternate cell death mechanism , mediated via ceramide synthase ( CS)-stimulated de novo synthesis of the proapoptotic sphingolipid ceramide , and p53-independent apoptosis of crypt SCCs . We previously reported that ataxia-telangiectasia mutated deficiency resets the primary radiation lethal pathway , allowing CS-mediated apoptosis at the low-dose range of radiation . The mechanism for this event , termed target reordering , remains unknown . Here , we show that inactivation of DNA damage repair pathways signals CS-mediated apoptosis in crypt SCCs , presumably via persistent unrepaired DNA double-strand breaks ( DSBs ) . Genetic loss of function of sensors and transducers of DNA DSB repair confers the CS-mediated lethal pathway in intestines of sv129/B6Mre11(ATLD1/ATLD1) and C57BL/6(Prkdc/SCID) ( severe combined immunodeficient ) mice exposed to low-dose radiation . In contrast , CS-mediated SCC lethality was mitigated in irradiated gain-of-function Rad50(s/s) mice , and epistasis studies order Rad50 upstream of Mre11 . These studies suggest unrepaired DNA DSBs as causative in target reordering in intestinal SCCs . As such , we provide an in vivo model of DNA damage repair that is standardized , can be exploited to understand allele-specific regulation in intact tissue , and is pharmacologically tractable . OUTPUT: genomic instability and mutation INPUT: The capacity to repair DNA damage is an important factor that affects the therapeutic outcome in cancer treatment . To clarify the cellular repair response , we investigated the kinetics of DNA excision repair initiated by 1,3-bis(2-chloroethyl)-1-nitrosourea ( BCNU ) in human leukemia CCRF-CEM cells at an exponential growth phase in vitro . Using the alkaline single-cell gel electrophoresis ( comet ) assay , we quantitated the repair kinetics as the amount of DNA single-strand breaks that were generated from the incision and were diminished by the rejoining in the repair process . CEM cells could initiate DNA excision repair in response to BCNU by starting an incision reaction . However , the incision capacity came to a plateau at a concentration of 80 to 100 microM or after an incubation time of 90 to 120 minutes . When the cells were pulsed with 40 microM BCNU , the maximal incision occurred at the end of the incubation period , and the repair process was completed within 4 hours When cells were treated with 100 microM BCNU , the incised DNA was not rejoined at 4 hours , suggesting that the repair was not completed . Higher concentrations might surpass the cellular capacity for repair and would be associated with increased cell death . Evaluation of the repair process may provide a clue for therapeutic strategies to improve clinical efficacy if accelerated DNA repair is responsible for the drug resistance . OUTPUT:

genomic instability and mutation;resisting cell death

HoC_dynamic_5_shot26

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: Genomic instability drives tumorigenesis , but how it is initiated in sporadic neoplasias is unknown . In early preneoplasias , alterations at chromosome fragile sites arise due to DNA replication stress . A frequent , perhaps earliest , genetic alteration in preneoplasias is deletion within the fragile FRA3B/FHIT locus , leading to loss of Fhit protein expression . Because common chromosome fragile sites are exquisitely sensitive to replication stress , it has been proposed that their clonal alterations in cancer cells are due to stress sensitivity rather than to a selective advantage imparted by loss of expression of fragile gene products . Here , we show in normal , transformed , and cancer-derived cell lines that Fhit-depletion causes replication stress-induced DNA double-strand breaks . Using DNA combing , we observed a defect in replication fork progression in Fhit-deficient cells that stemmed primarily from fork stalling and collapse . The likely mechanism for the role of Fhit in replication fork progression is through regulation of Thymidine kinase 1 expression and thymidine triphosphate pool levels ; notably , restoration of nucleotide balance rescued DNA replication defects and suppressed DNA breakage in Fhit-deficient cells . Depletion of Fhit did not activate the DNA damage response nor cause cell cycle arrest , allowing continued cell proliferation and ongoing chromosomal instability . This finding was in accord with in vivo studies , as Fhit knockout mouse tissue showed no evidence of cell cycle arrest or senescence yet exhibited numerous somatic DNA copy number aberrations at replication stress-sensitive loci . Furthermore , cells established from Fhit knockout tissue showed rapid immortalization and selection of DNA deletions and amplifications , including amplification of the Mdm2 gene , suggesting that Fhit loss-induced genome instability facilitates transformation . We propose that loss of Fhit expression in precancerous lesions is the first step in the initiation of genomic instability , linking alterations at common fragile sites to the origin of genome instability . OUTPUT: genomic instability and mutation;sustaining proliferative signaling;enabling replicative immortality INPUT: Genomic instability has long been recognized as the main feature of neoplasia and a factor modulating individual cancer susceptibility . There are attempts to find effective assays of both individual DNA repair capacity and genetic instability , and their relation to the cancer risk . Genetic predisposition plays an important role in the etiology and development of head and neck squamous cell carcinoma ( HNSCC ) . The aim of our study was to search for a correlation between chromosomal instability and DNA repair capacity in HNSCC patients and healthy controls . The chromosomal instability was measured by the number of bleomycin ( BLM)-induced chromosomal aberrations and diepoxybutane ( DEB)-induced sister chromatid exchanges . The DNA repair capacity was assessed using the DEB-induced adaptive response ( AR ) . The HNSCC patients in our study showed a significant increase in chromosomal instability after a preterminal exposure of their lymphocytes to either BLM for the last 5 h or DEB for the last 24 h of incubation . However , the AR was higher in HNSCC patients than in the control group , suggesting an increase in the DNA repair capacity in the cancer patients as compared to the control . There is no correlation between the DNA repair capacity estimated on the basis of preterminal exposures to BLM and DEB and the DNA repair capacity estimated on the basis of the adaptive response to DEB . The preterminal exposure and the adaptive response test may activate different DNA repair mechanisms . OUTPUT: genomic instability and mutation INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: Class switch recombination ( CSR ) in B lymphocytes is initiated by introduction of multiple DNA double-strand breaks ( DSBs ) into switch ( S ) regions that flank immunoglobulin heavy chain ( IgH ) constant region exons . CSR is completed by joining a DSB in the donor S mu to a DSB in a downstream acceptor S region ( e.g. , S gamma1 ) by end-joining . In normal cells , many CSR junctions are mediated by classical nonhomologous end-joining ( C-NHEJ ) , which employs the Ku70/80 complex for DSB recognition and XRCC4/DNA ligase 4 for ligation . Alternative end-joining ( A-EJ ) mediates CSR , at reduced levels , in the absence of C-NHEJ , even in combined absence of Ku70 and ligase 4 , demonstrating an A-EJ pathway totally distinct from C-NHEJ . Multiple DSBs are introduced into S mu during CSR , with some being rejoined or joined to each other to generate internal switch deletions ( ISDs ) . In addition , S-region DSBs can be joined to other chromosomes to generate translocations , the level of which is increased by absence of a single C-NHEJ component ( e.g. , XRCC4 ) . We asked whether ISD and S-region translocations occur in the complete absence of C-NHEJ ( e.g. , in Ku70/ligase 4 double-deficient B cells ) . We found , unexpectedly , that B-cell activation for CSR generates substantial ISD in both S mu and S gamma1 and that ISD in both is greatly increased by the absence of C-NHEJ . IgH chromosomal translocations to the c-myc oncogene also are augmented in the combined absence of Ku70 and ligase 4 . We discuss the implications of these findings for A-EJ in normal and abnormal DSB repair . OUTPUT: genomic instability and mutation INPUT: Transfected linear DNA molecules are substrates for double-strand break ( DSB ) repair in mammalian cells . The DSB repair process can involve recombination between the transfected DNA molecules , between the transfected molecules and chromosomal DNA , or both . In order to determine whether these different types of repair events are linked , we devised assays enabling us to follow the fate of linear extrachromosomal DNA molecules involved in both interplasmid and chromosome-plasmid recombination , in the presence or absence of a pre-defined chromosomal DSB . Plasmid-based vectors were designed that could either recombine via interplasmid recombination or chromosome-plasmid recombination to produce a functional beta-galactosidase ( betagal ) fusion gene . By measuring the frequency of betagal+ cells at 36 h post-transfection versus the frequency of betagal+ clones after 14 days , we found that the number of cells containing extrachromosomal recombinant DNA molecules at 36 h ( i.e. , betagal+ ) , either through interplasmid or chromosome-plasmid recombination , was nearly the same as the number of cells integrating these recombinant molecules . Furthermore , when a predefined DSB was created at a chromosomal site , the extrachromosomal recombinant DNA molecules were shown to integrate preferentially at that site by Southern and fiber-FISH ( fluorescence in situ hybridization ) analysis . Together these data indicate that the initial recombination event can potentiate or commit extrachromosomal DNA to integration in the genome at the site of a chromosomal DSB . The efficiency at which extrachromosomal recombinant molecules are used as substrates in chromosomal DSB repair suggests extrachromosomal DSB repair can be coupled to the repair of chromosomal DSBs in mammalian cells . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot27

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Blockage of the metastasis process remains a significant clinical challenge , requiring innovative therapeutic approaches . For this purpose , molecules that inhibit matrix metalloproteinases activity or induce the expression of their natural inhibitor , the tissue inhibitor of metalloproteinases ( TIMPs ) , are potentially interesting . In a previous study , we have shown that synthetic ligands binding to cell surface nucleolin/nucleophosmin and known as HB 19 for the lead compound and NucAnt 6L ( N6L ) for the most potent analog , inhibit both tumor growth and angiogenesis . Furthermore , they prevent metastasis in a RET transgenic mice model which develops melanoma . Here , we investigated the effect of N6L on the invasion capacity of MDA-MB-435 melanoma cells . Our results show that the multivalent pseudopeptide N6L inhibited Matrigel invasion of MDA-MB-435 cells in a modified Boyden chamber model . This was associated with an increase in TIMP-3 in the cell culture medium without a change in TIMP-3 mRNA expression suggesting its release from cell surface and/or extracellular matrix . This may be explained by our demonstrated N6L interaction with sulfated glycosaminoglycans and consequently the controlled bioavailability of glycosaminoglycan-bound TIMP-3 . The implication of TIMP-3 in N6L-induced inhibition of cell invasion was evidenced by siRNA silencing experiments showing that the loss of TIMP-3 expression abrogated the effect of N6L . The inhibition of tumor cell invasion by N6L demonstrated in this study , in addition to its previously established inhibitory effect on tumor growth and angiogenesis , suggests that N6L represents a promising anticancer drug candidate warranting further investigation . OUTPUT: activating invasion and metastasis;inducing angiogenesis INPUT: Multiple myeloma is characterized by the clonal expansion of malignant plasma cells ( multiple myeloma cells [ MMCs] ) , in the bone marrow . Osteolytic bone lesions are detected in 80% of patients because of increased osteoclastic bone resorption and reduced osteoblastic bone formation . MMCs are found closely associated with sites of increased bone resorption . Osteoclasts strongly support MMC survival in vitro . To further elucidate the mechanisms involved in osteoclast/MMC interaction , we have identified 552 genes overexpressed in osteoclasts compared with other bone marrow cell subpopulations . Osteoclasts express specifically genes coding for 4 CCR2-targeting chemokines and genes coding for MMC growth factors . An anti-CCR2 monoclonal antibody blocked osteoclast chemoattractant activity for MMC , and CCR2 chemokines are also MMC growth factors , promoting mitogen-activated protein kinase activation in MMC . An anti-insulin growth factor-1 receptor monoclonal antibody completely blocked the osteoclast-induced survival of MMC suppressing both osteoclast and MMC survival . Specific a proliferation-inducing ligand or IL-6 inhibitors partially blocked osteoclast-induced MMC survival . These data may explain why newly diagnosed patients whose MMC express high levels of CCR2 present numerous bone lesions . This study displays additional mechanisms involved in osteoclast/MMC interaction and suggests using CCR2 and/or insulin growth factor-1 targeting strategies to block this interaction and prevent drug resistance . OUTPUT: sustaining proliferative signaling INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: INTRODUCTION Necrosis at the tumor center is a common feature of aggressive breast cancers and has been associated with poor prognosis . It is commonly identified by means of invasive histopathology , which often correlates with morbidity and potential tumor cell dissemination , and limits the reconstruction of the whole necrotic domain . In this study we hypothesized that non covalent association to serum albumin ( SA ) and covalent binding to ligands for tumor-abundant cell receptors should synergistically drive selective accumulation and prolonged retention of imaging and therapeutic agents in breast tumor necrotic domains enabling in vivo identification , imaging and possibly treatment of such tumors . METHODS Cyclo-Arg-Gly-Asp-D-Phe-Lys ( c(RGDfK) ) were conjugated to bacteriochlorophyll-derivatives ( Bchl-Ds ) , previously developed as photodynamic agents , fluorescent probes and metal chelators in our lab . The c(RGDfK) component drives ligation to alphaVbeta3 integrin receptors over-expressed by tumor cells and neo-vessels , and the Bchl-D component associates to SA in a non-covalent manner . STL-6014 , a c(RGDfK)-Bchl-D representative , was i.v. injected to CD-1 , nude female mice bearing necrotic and non-necrotic human MDA-MB-231-RFP breast cancer tumors . The fluorescence signals of the Bchl-Ds and RFP were monitored over days after treatment , by quantitative whole body imaging and excised tumor/tissue samples derived thereof . Complementary experiments included competitive inhibition of STL-6014 uptake by free c(RGDfK) , comparative pharmacokinetics of nonconjugated c(RGDfK) Bchl-D ( STL-7012 ) and of two human serum albumin ( HSA ) conjugates : HSA-STL-7012 and HSA-STL-6014 . RESULTS STL-6014 and STL-7012 formed complexes with HSA ( HSA/STL-6014 , HSA/STL-7012 ) . STL-6014 , HSA-STL-7012 and HSA-STL-6014 , selectively accumulated at similar rates , in tumor viable regions over the first 8 h post administration . They then migrated into the necrotic tumor domain and presented tumor half lifetimes ( T1/2 ) in the range of days where T1/2 for HSA-STL-6014 &gt ; STL-6014 &gt ; HSA-STL-7012 . No accumulation of STL-7012 was observed . Pre-injection of c(RGDfK) excess , prevented the uptake of STL-6014 in the small , but not in the large tumors . CONCLUSIONS Non-covalent association to SA and covalent binding to c(RGDfK) , synergistically enable the accumulation and prolonged retention of Bchl-Ds in the necrotic regions of tumors . These findings provide novel guidelines and strategy for imaging and treatment of necrotic tumors . OUTPUT: resisting cell death INPUT: BACKGROUND The presence of distant metastases from colorectal cancer ( CRC ) does not preclude curative treatment . Early detection of pulmonary metastases at a potentially curable stage could improve survival . The aim of the present study was to assess the prognostic significance of commonly reported clinicopathologic features to identify high-risk patients who would likely benefit from more intensive chest surveillance for pulmonary metastases . MATERIAL AND METHOD A total of 351 consecutive patients , with surgical stages I-III colorectal cancer , who underwent curative resection at Phramongkutklao hospital from 1999 to 2005 , were followed regularly according to the established guidelines with routine physical examination , serum carcinoembryonic antigen ( CEA ) and colonoscopic surveillance . Imaging studies for detecting metastases were computed tomography ( CT ) , plain film radiography , and ultrasonograpy . Clinical and pathologic features were analyzed for their association with pulmonary metastasis . RESULTS There were 145 patients who had been operated for longer than five years after curative intent surgery . Of these , nineteen patients were lost to follow-up or died from other causes that were unrelated to colorectal cancer . Pulmonary metastases were detected in 26 patients by either CXR or CT scan . Median time to pulmonary metastasis was 19 months ( 95 percent CI , 12-35 ) . According to an univariate analysis , with log-rank test , identified four factors associated with pulmonary metastasis : Tumor stage T4 , Nodal stage N2 , elevation of serum CEA &gt ; 3.4 ng/ml and presence of lymphovascular invasion(LVI) . According to a multivariate analysis , with Cox regression , found an elevation of serum CEA &gt ; 3.4 ng/ml which was an independent factor that was significantly associated with pulmonary metastasis ( Hazard ratio ( HR ) , 8.9 ; 95 percent CI , 3.6-22 ; p &lt ; 0.01 ) . The present study revealed that 50 percent of patients who had more than one of these risk factors would eventually develop pulmonary metastases . CONCLUSION An elevation of serum CEA &gt ; or = 3.4 ng/ml was found as an independent factor that was significantly associated with pulmonary metastasis whereas tumor stage T4 , nodal stage N2 and presence of lymphovascular invasion ( LVI ) were not independent clinicopathologic features associated with subsequent pulmonary metastases . Chest CT scan has greater sensitivity than chest radiography in detection of pulmonary metastasis and should be considered as an imaging study of choice for intensive chest surveillance for patients who had more than one of these risk factors . OUTPUT: activating invasion and metastasis INPUT: Tumor metastasis represents a complex multistep process that requires migration , invasion , and angiogenesis . In this study , we examined the impact of molecular blockade of the epidermal growth factor receptor on the invasive and metastatic capacity of human squamous cell carcinoma ( SCC ) of the head and neck using in vitro and in vivo model systems . Treatment with the anti-epidermal growth factor receptor antibody C225 attenuated the migration of SCC-1 tumor cells through a chemotaxis chamber in a dose-dependent manner . Incubation of SCC cells with 10-100 nM C225 for 4 h resulted in 40-60% inhibition of cell migration . Furthermore , in the presence of C225 , the capacity of SCC-1 to invade across a layer of extracellular matrix ( Matrigel ) was significantly inhibited . Using an in vivo orthotopic floor-of-mouth xenograft model , locoregional tumor invasion of SCC-1 into muscle , vessel , bone , and perineural tissues was inhibited in C225-treated mice . This inhibition was additionally characterized by down-regulation in the expression of matrix metalloproteinase-9 . These data suggest that inhibition of metastatic potential by C225 may be mediated via decreased migration and invasion of SCC cells . Regarding angiogenesis in vitro , we first studied human umbilical vascular endothelial cells , which established a capillary-like network structure ( tube formation ) in the presence of reconstituted Matrigel . Treatment with C225 reduced cell-to-cell interaction of human umbilical vascular endothelial cells , resulting in disruption of tube formation . The effect of C225 was additionally examined using an in vivo tumor xenograft neovascularization model of angiogenesis . Systemic treatment with C225 not only reduced tumor growth and the number of blood capillaries but also hindered the growth of established vessels toward the tumor . Taken together , these results provide evidence that C225 can suppress tumor-induced neovascularization and metastasis in SCC of the head and neck . OUTPUT:

activating invasion and metastasis;inducing angiogenesis

HoC_dynamic_5_shot28

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: Comparative mutagenesis studies of N-(2'-deoxyguanosin-8-yl)-2-acetylaminofluorene ( dG-AAF ) and N-(2'-deoxyguanosin-8-yl)-2-aminofluorene ( dG-AF ) adducts positioned in the Nar I restriction enzyme site were performed using Escherichia coli ( E. coli ) and simian kidney ( COS-7 ) cells . Oligodeoxynucleotides ( (5)(')TCCTCG(1)G(2)CG(3)CCTCTC ) containing a recognition sequence for the Nar I restriction enzyme were modified site-specifically with dG-AAF or dG-AF . Modified and unmodified oligomers inserted into single-stranded phagemid shuttle vectors were used to transform E. coli or to transfect COS-7 cells . Following replication in host cells , progeny plasmids were recovered and analyzed for mutations . In SOS-induced E. coli , dG-AAF primarily induced one- and two-base deletions . The mutational frequency varied , depending on the position modified in the Nar I site ; 91% two-base deletions were observed at G(3) , while 8.4% and 2.8% deletions were detected at G(2) and G(1) , respectively . In contrast , dG-AF at any position in the Nar I site failed to produce deletions , generating primarily G --&gt ; T transversions ( mutational frequency , 7.6-8.4% ) . In COS-7 cells , both dG-AAF and dG-AF primarily induced G --&gt ; T transversions . Mutation frequencies for dG-AAF were 9.4-24% , the highest values being at G(1) and G(3) . Mutation frequencies for dG-AF were 9.3-21% , the higher value at G(2) . We conclude from this study that the mutation potential of dG-AAF and dG-AF depends on the structure of the adduct , the sequence context of the lesion , and the host cell used for the experiment . OUTPUT: genomic instability and mutation INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: The use of substrates containing well defined adducts at precise sites , is required to perform a careful analysis of the toxic and mutagenic potential of a lesion . As a first step in this direction the octamer 5'-d(CCGGCGGT) , containing the sequence of the codons 12 d(GGC) and 13 d(GGT) of the human H-ras gene , was reacted with the antitumoral drug cis-diamminedichloroplatinum(II) . The platinated products have been purified by HPLC . A first set of experiments , including enzymatic digestions with nuclease P1 followed by alkaline phosphatase and acid-catalysed hydrolysis , allowed us to determine which bases were engaged in the cis-DDP lesions . Our results indicate that only guanine residues were chelated with cisplatin to yield bifunctional adducts . Furthermore , by performing enzymatic digestions with phosphodiesterases , we have located the adducts with respect to the 5 ' end of the octamer . Among the purified and characterized platinated oligonucleotides , three present a particular interest , since we have shown here that the cis-d(GpG) adduct is precisely situated either at the d(GGC) or at the d(GGT) or at both sites of their sequence . OUTPUT: genomic instability and mutation INPUT: "The main objective of the present investigation was to study the urinary neopterin excretion in the context of the activation of the adaptive cellular immune system at the tumor site . For this purpose , we compared pre-treatment urinary neopterin levels measured in 92 ovarian cancer patients , with intratumoral levels of mRNA transcripts from factors either involved in the adaptive antitumor immune defense ( CD3 , IFN-γ , IRF-1 , IRF-2 , SOCS1 and iNOS ) or immune tolerance ( FoxP3 ) . This study did not reveal an association between urinary neopterin and one of these investigated "" on tumor site transcripts "" . From all the factors reflecting the magnitude of the local adaptive antitumor response , intratumoral IRF-1 expression above the edge of the 25th percentile was found to predict most reliably favorable progression-free ( median 34 months vs. 10 months ; p &lt ; 0.001 ) and overall ( median 52 months vs. 16 months ; p &lt ; 0.001 ) survival . In contrast , pre-treatment urinary neopterin excretion above 275 μmol/mol creatinine , which indicates an unspecific activation of the innate immune system , was associated with a very poor overall survival with a median of only 11 months when compared with a median overall survival of 40 months in patients with lower urinary neopterin excretion ( p = 0.021 ) . Interestingly , the considerable survival benefit in patients with high IRF-1-expressing cancers was completely abrogated as well for progression-free as for overall survival when urinary neopterin concentrations were found to be concomitantly elevated . These findings demonstrate that in ovarian carcinomas the unspecific "" cancer-related inflammation "" contributes to a significant subversion of the adaptive antitumor immune defense mounted at the tumor site ." OUTPUT: tumor promoting inflammation INPUT: Although there have been numerous studies of site-specific mutagenesis by dGuo adducts of benzo[a]pyrene diol epoxides ( B[a]P DEs ) , the present study represents the first example of site-specific mutagenesis by dGuo adducts of the highly carcinogenic benzo[c]phenanthrene 3,4-diol 1,2-epoxides ( B[c]Ph DEs ) . The eight adducts that would result from cis- and trans-opening at C-1 of four optically active isomers of B[c]Ph DEs by the N(2)-amino group of dGuo were incorporated into 5'-TTCGAATCCTTCCCCC ( context III ) and 5'-GGGGTTCCCGAGCGGC ( context IV ) at the underlined site . These modified oligonucleotides along with unmodified controls were ligated into single-stranded M13mp7L2 , which were then used to transfect SOS-induced Escherichia coli . Upon replication of the lesions in each of the two sequence contexts , mutational analysis of the progeny was performed by differential hybridization . For the 16 adducts , the mutation frequencies varied over 2 orders of magnitude with a reasonably even distribution ( 0.4-1% for three adducts , 1-2% for six adducts , 3-7.4% for five adducts , and one adduct each at 11 and 39% ) . For all but this last adduct , the mutation frequency for a given B[c]Ph DE adduct was less than for its B[a]P analogue with the same stereochemistry in the same sequence . For the vectors containing adducts with S configuration at the site of attachment of the hydrocarbon to the dGuo base , the main base substitution was G --&gt ; T followed by G --&gt ; A. In contrast , for the vectors containing adducts with R configuration , the main base substitution was G --&gt ; A. The most notable observation in the present study is the low frequency of mutations induced by the B[c]Ph DE-dGuo adducts relative to their B[a]P counterparts . A possible structural basis for this difference is proposed . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot29

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Acrolein ( Acr ) is a ubiquitous environmental pollutant found in cigarette smoke and automobile exhaust . It can also be produced endogenously by oxidation of polyunsaturated fatty acids . The Acr-derived 1,N(2)-propanodeoxyguanosine ( Acr-dG ) adducts in DNA are mutagenic lesions that are potentially involved in human cancers . In this study , monoclonal antibodies were raised against Acr-dG adducts and characterized using ELISA . They showed strong reactivity and specificity toward Acr-dG , weaker reactivity toward crotonaldehyde- and trans-4-hydroxy-2-nonenal-derived 1,N(2)-propanodeoxyguanosines , and weak or no reactivity toward 1,N(6)-ethenodeoxyadenosine and 8-oxo-deoxyguanosine . Using these antibodies , we developed assays to detect Acr-dG in vivo : first , a simple and quick FACS-based assay for detecting these adducts directly in cells ; second , a highly sensitive direct ELISA assay for measuring Acr-dG in cells and tissues using only 1 μg of DNA without DNA digestion and sample enrichment ; and third , a competitive ELISA for better quantitative measurement of Acr-dG levels in DNA samples . The assays were validated using Acr-treated HT29 cell DNA samples or calf thymus DNA , and the results were confirmed by LC-MS/MS-MRM . An immunohistochemical assay was also developed to detect and visualize Acr-dG in HT29 cells as well as in human oral cells . These antibody-based methods provide useful tools for the studies of Acr-dG as a cancer biomarker and of the molecular mechanisms by which cells respond to Acr-dG as a ubiquitous DNA lesion . OUTPUT: genomic instability and mutation INPUT: We report the formation , detection , quantitation and structural characterization of products resulting from the adduction of deoxynucleosides ( deoxyadenosine , deoxyguanosine , deoxycytidine and 5-methyldeoxycytidine ) to the catechol estrogens ( CE ) of estrone , estradiol-17beta and estradiol-17 alpha . The crude products are obtained in a one-pot synthesis through oxidation of catechols to quinones and subsequent Michael-type reaction with the deoxynucleosides in acidic medium . In all experiments , adducts are detected by electrospray ionization mass spectrometry analysis after HPLC separation ( LC/ESI/MS(n) ) . The two pyrimidines deoxycytidine and 5-methyldeoxycytidine yield only CE adducts to deoxynucleosides , which correspond to stable adducts on DNA . For purines , the results depend on the CE ( 2,3- or 3,4-catechols ) used , the function and configuration on carbon 17 ( ketone for estrone , alcohol for alpha and beta isomers of estradiol ) , and on the purine itself ( deoxyadenosine or deoxyguanosine ) . Both stable adducts and deglycosylated adducts are formed , and therefore formation of stable adducts on DNA as well as the loss of purines from the DNA strands could be possible . MS(2) and MS(3) experiments prove to be relevant for further structural determinations , enabling in some cases the elucidation of the regiochemistry of adduction on the A and B rings of the steroid moiety . OUTPUT: genomic instability and mutation INPUT: The ALKBH family of proteins are highly expressed in various types of human cancer where they are involved in tumor growth and progression . However , multiple isoforms of ALKBH exist and the effect of individual isoforms on the development of urinary bladder cancer is unknown , particularly the molecular mechanisms involved in the progression from a noninvasive to invasive phenotype . We examined the role and function of ALKBH2 in human bladder cancer development in vitro and provide the first report that suppression of ALKBH2 in a human urothelial carcinoma cell line , KU7 , reduced the expression of the transmembrane mucin protein , MUC1 , and induced G1 cell cycle arrest . Moreover , reduction of ALKBH2 suppressed epithelial to mesenchymal transition ( EMT ) via increasing E-cadherin and decreasing vimentin expression . Transfection of MUC1 siRNA inhibited cell proliferation and EMT to the same extent as ALKBH2 gene silencing in vitro . ALKBH2 knockdown significantly suppressed MUC1 expression and tumor volume of bladder cancers in vivo as assessed in an orthotopic mouse model using ALKBH2 shRNA transfected KU7 cells . Immunohistochemical examination showed high expression levels of ALKBH2 in human urothelial carcinoma samples , especially in high-grade , superficially and deeply invasive carcinomas ( pT(1) and >pT(2) ) , and in carcinoma in situ but not in normal urothelium . This study demonstrates that ALKBH2 is an upstream molecule of the oncoprotein , MUC1 , and regulates cell cycle and EMT , resulting in progression of urothelial carcinomas . OUTPUT: evading growth suppressors;sustaining proliferative signaling;activating invasion and metastasis INPUT: Extensive biochemical and structural analyses have been performed on the putative DNA repair proteins of hyperthermophilic archaea , in contrast to the few genetic analyses of the genes encoding these proteins . Accordingly , little is known about the repair pathways used by archaeal cells at high temperature . Here , we attempted to disrupt the genes encoding the potential repair proteins in the genome of the hyperthermophilic archaeon Thermococcus kodakaraensis . We succeeded in isolating null mutants of the hjc , hef , hjm , xpb , and xpd genes , but not the radA , rad50 , mre11 , herA , nurA , and xpg/fen1 genes . Phenotypic analyses of the gene-disrupted strains showed that the xpb and xpd null mutants are only slightly sensitive to ultraviolet ( UV ) irradiation , methyl methanesulfonate ( MMS ) and mitomycin C ( MMC ) , as compared with the wild-type strain . The hjm null mutant showed sensitivity specifically to mitomycin C. On the other hand , the null mutants of the hjc gene lacked increasing sensitivity to any type of DNA damage . The Hef protein is particularly important for maintaining genome homeostasis , by functioning in the repair of a wide variety of DNA damage in T. kodakaraensis cells . Deletion of the entire hef gene or of the segments encoding either its nuclease or helicase domain produced similar phenotypes . The high sensitivity of the Δhef mutants to MMC suggests that Hef performs a critical function in the repair process of DNA interstrand cross-links . These damage-sensitivity profiles suggest that the archaeal DNA repair system has processes depending on repair-related proteins different from those of eukaryotic and bacterial DNA repair systems using homologous repair proteins analyzed here . OUTPUT: genomic instability and mutation INPUT: DNA-protein cross-links ( DPCs ) are formed upon exposure to a variety of chemical and physical agents and pose a threat to genomic integrity . In particular , acrolein and related aldehydes produce DPCs , although the chemical linkages for such cross-links have not been identified . Here , we report that oligodeoxynucleotides containing 1,N(2)-deoxyguanosine adducts of acrolein , crotonaldehyde , and trans-4-hydroxynonenal can form cross-links with the tetrapeptide Lys-Trp-Lys-Lys . We concluded that complex formation is mediated by a Schiff base linkage because DNA-peptide complexes were covalently trapped following reduction with sodium cyanoborohydride , and pre-reduction of adducted DNAs inhibited complex formation . A previous NMR study demonstrated that duplex DNA catalyzes ring opening for the acrolein-derived gamma-hydroxy-1,N(2)-propanodeoxyguanosine adduct to yield an aldehydic function ( de los Santos , C. , Zaliznyak , T. , and Johnson , F . ( 2001 ) J. Biol . Chem. 276 , 9077-9082 ) . Consistent with this earlier observation , the adducts under investigation were more reactive in duplex DNA than in single-stranded DNA , and we concluded that the ring-open aldehydic moiety is the induced tautomer in duplex DNA for adducts exhibiting high relative reactivity . Adducted DNA cross-linked to Arg-Trp-Arg-Arg and Lys-Trp-Lys-Lys with comparable efficiency , and N(alpha)-acetylation of peptides dramatically inhibited trapping ; thus , the reactive nucleophile is located at the N-terminal alpha-amine of the peptide . These data suggest that Schiff base chemistry can mediate DPC formation in vivo following the formation of stable aldehyde-derived DNA adducts . OUTPUT: genomic instability and mutation INPUT: The Escherichia coli AlkB protein protects against the cytotoxicity of methylating agents by repair of the DNA lesions 1-methyladenine and 3-methylcytosine , which are generated in single-stranded stretches of DNA . AlkB is an alpha-ketoglutarate- and Fe(II)-dependent dioxygenase that oxidizes the relevant methyl groups and releases them as formaldehyde . Here , we identify two human AlkB homologs , ABH2 and ABH3 , by sequence and fold similarity , functional assays , and complementation of the E. coli alkB mutant phenotype . The levels of their mRNAs do not appear to correlate with cell proliferation but tissue distributions are different . Both enzymes remove 1-methyladenine and 3-methylcytosine from methylated polynucleotides in an alpha-ketoglutarate-dependent reaction , and act by direct damage reversal with the regeneration of the unsubstituted bases . AlkB , ABH2 , and ABH3 can also repair 1-ethyladenine residues in DNA with the release of acetaldehyde . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot30

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: DNA mismatch repair ( MMR ) of simple base mismatches and small insertion-deletion loops in eukaryotes is initiated by the binding of the MutS homolog 2 ( MSH2)-MSH6 heterodimer to mismatched DNA . Cadmium ( Cd ) is a genotoxic heavy metal that has been recognized as a human carcinogen . Oxidant stress and inhibition of DNA repair have been proposed as major factors underlying Cd genotoxicity . Our previous studies indicated the ability of Cd to disturb the gene expression of MSH6 in zebrafish ( Danio rerio ) embryos . This study was undertaken to explore if Cd-induced oxidative stress down-regulated MSH gene activities . Following the exposure of zebrafish embryos at 1 h post fertilization ( hpf ) to sublethal concentrations of Cd at 3-5 μM for 4 or 9 h , a parallel down-regulation of MSH2 , MSH6 and Cu/Zn superoxide dismutase ( Cu/Zn-SOD ) gene expression was detected by real-time RT-PCR and the expression levels were 40-50% of control after a 9-h exposure . Cd exposure also induced oxidative stress , yet no inhibition of catalase gene activity was observed . Whole mount in situ hybridization revealed a wide distribution of msh6 mRNA in the head regions of 10 hpf embryos and pretreatment of embryos with antioxidants butylhydroxytoluene ( BHT ) , d-mannitol or N-acetylcysteine ( NAC ) at 1-10 μM restored Cd-suppressed msh6 expression . QPCR confirmed the protective effects of antioxidants on Cd-suppressed msh2/msh6 mRNA production . Down-regulated MSH gene activities reaching about 50% of control were also induced in embryos exposed to paraquat , a reactive oxygen species ( ROS)-generating herbicide , or hydrogen peroxide at 200 μM . Hence , Cd at sublethal levels down-regulates msh2/msh6 expression primarily via ROS as signaling molecules . The transcriptional activation of human msh6 is known to be fully dependent on the specificity factor 1 ( Sp1 ) . Cd failed to inhibit the DNA binding activity of zebrafish Sp1 unless at lethal concentrations based on band shift assay , therefore excluding the involvement of Sp1 inactivation in Cd-induced MSH gene inhibition in zebrafish embryos . OUTPUT: tumor promoting inflammation INPUT: DNA mismatch repair ( MMR ) is the process by which incorrectly paired DNA nucleotides are recognized and repaired . A germline mutation in one of the genes involved in the process may be responsible for a dominantly inherited cancer syndrome , hereditary nonpolyposis colon cancer . Cancer progression in predisposed individuals results from the somatic inactivation of the normal copy of the MMR gene , leading to a mutator phenotype affecting preferentially repeat sequences ( microsatellite instability , MSI ) . Recently , we identified children with a constitutional deficiency of MMR activity attributable to a mutation in the h MLH1 gene . These children exhibited a constitutional genetic instability associated with clinical features of de novo neurofibromatosis type 1 ( NF1 ) and early onset of extracolonic cancer . Based on these observations , we hypothesized that somatic NF1 gene mutation was a frequent and possibly early event in MMR-deficient cells . To test this hypothesis , we screened for NF1 mutations in cancer cells . Genetic alterations were identified in five out of ten tumor cell lines with MSI , whereas five MMR-proficient tumor cell lines expressed a wild-type NF1 gene . Somatic NF1 mutations were also detected in two primary tumors exhibiting an MSI phenotype . Finally , a 35-bp deletion in the murine Nf1 coding region was identified in mlh1-/- mouse embryonic fibroblasts . These observations demonstrate that the NF1 gene is a mutational target of MMR deficiency and suggest that its inactivation is an important step of the malignant progression of MMR-deficient cells . OUTPUT: genomic instability and mutation INPUT: The DNA mismatch repair ( MMR ) pathway corrects specific types of DNA replication errors that affect microsatellites and thus is critical for maintaining genomic integrity . The genes of the MMR pathway are highly conserved across different organisms . Likewise , defective MMR function universally results in microsatellite instability ( MSI ) which is a hallmark of certain types of cancer associated with the Mendelian disorder hereditary nonpolyposis colorectal cancer . ( Lynch syndrome ) . To identify previously unrecognized deleted genes or loci that can lead to MSI , we developed a functional genomics screen utilizing a plasmid containing a microsatellite sequence that is a host spot for MSI mutations and the comprehensive homozygous diploid deletion mutant resource for Saccharomyces cerevisiae . This pool represents a collection of non-essential homozygous yeast diploid ( 2N ) mutants in which there are deletions for over four thousand yeast open reading frames ( ORFs ) . From our screen , we identified a deletion mutant strain of the PAU24 gene that leads to MSI . In a series of validation experiments , we determined that this PAU24 mutant strain had an increased MSI-specific mutation rate in comparison to the original background wildtype strain , other deletion mutants and comparable to a MMR mutant involving the MLH1 gene . Likewise , in yeast strains with a deletion of PAU24 , we identified specific de novo indel mutations that occurred within the targeted microsatellite used for this screen . OUTPUT: genomic instability and mutation INPUT: Mice defective in the mismatch repair ( MMR ) gene Msh2 manifest an enhanced predisposition to skin cancer associated with exposure to UVB radiation . This predisposition is further heightened if the mice are additionally defective for the nucleotide excision repair gene Xpc . To test the hypothesis that the predisposition of Msh2 mutant mice to skin cancer reflects a mutator phenotype associated with increased proliferation of skin cells following exposure to UV radiation , Msh2 mutant mice were exposed to the tumor promoter TPA . Such mice showed a robust proliferative response in the skin , but did not manifest evidence of dysplasia or neoplasia . We conclude that the predisposition of Msh2 mice to UVB radiation-induced skin cancer reflects an interaction between the processes of mismatch repair and some other excision repair mode , the exact nature of which remains to be established . OUTPUT: genomic instability and mutation INPUT: The β2-adrenergic receptor ( β2AR ) mediates the effects of chronic stress in several neoplasms , however , β2AR signaling is impaired by hypoxia in various tissues . While hypoxia is a common feature significant in the progression of solid tumors , little is known about the effect of hypoxia on β2AR signaling in the tumor microenvironment . Previously , it has been reported that the systemic administration of mesenchymal stem cells ( MSCs ) increased the engraftment and metastatic colonization of rat osteosarcoma ( OS ) cells . In the current study , the effect of MSCs on the hypoxia-induced desensitization of the β2AR in OS cells was investigated . Epinephrine , norepinephrine and isoproterenol increased the cellular proliferation of the rat OS cell line COS1NR and rat MSCs in a dose-dependent and β2AR antagonist-sensitive manner . While isoproterenol had significant proliferative effects on MSCs under normoxic and hypoxic conditions , COS1NR cells did not respond under hypoxic conditions . A sensitivity assay for the β2AR revealed that hypoxia impaired the sensitivity of COS1NR cells , whereas hypoxia did not affect MSCs . An immunoassay revealed no significant change in the expression of hypoxia-inducible factor-1α ( HIF1α ) in COS1NR cells , whilst an immunoassay demonstrated a 15% increase in MSCs following isoproterenol stimulation . In COS1NR cells co-cultured with MSCs under hypoxic conditions , isoproterenol caused a significant increase in proliferation and this effect was inhibited by an anti-interleukin ( IL)-6 antibody . A tumor formation assay in syngeneic rats revealed that the systemic administration of MSCs enhances the growth of OS and the effect of MSCs was inhibited by IL-6 neutralization . In conclusion , MSCs are resistant to the hypoxia-induced desensitization to β2AR . Hypoxia caused a siginificant desensitization of the β2AR in COS1NR cells alone , whereas MSCs may support tumor progression through cellular interactions . OUTPUT: sustaining proliferative signaling INPUT: The Msh2 DNA mismatch repair gene is one of five genes implicated in the pathogenesis of hereditary nonpolyposis colorectal cancer ( HNPCC ) . To address the possible mechanisms of the site-specific occurrence of HNPCC , the effect of Msh2 deficiency on mutations in different parts of the colon was investigated using the BC-1(lacI)/Msh2 double transgenic mouse . Compared to the Msh2(+/+) mice , Msh2(-/-) mice had an 8-9-fold increase of mutation frequency ( MF ) in the lacI gene from the cecum and the proximal and distal colon . The mutational spectra were also significantly different between Msh2(+/+) and Msh2(-/-) mice , with a significant increase in the frequency of -1 frameshifts and G:C-->A:T base substitutions in the repair-deficient mice . However , in spite of the site-specific predisposition of HNPCC in humans , we found no significant difference in the MF or mutation spectrum between the three parts of the colon in Msh2(+/+) , Msh2(+/-) , or Msh2(-/-) mice . In addition , 11 independent mutants harboring complex mutations within the lacI gene were recovered in the Msh2(-/-) mice . Interestingly , while the Msh2(+/-) mice displayed an overall MF similar to that observed in the wild-type mice , sequencing revealed a significantly different mutational spectrum between Msh2(+/+) and Msh2(+/-) mice , mainly characterized by an increase in -1 frameshifts . Due to the prevalence of frameshift mutations in HNPCC patients , this haploinsufficiency effect of the Msh2 gene in safeguarding genomic integrity may have important implications for human carrier status . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot31

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: A prominent feature of inflammatory diseases is endothelial dysfunction . Factors associated with endothelial dysfunction include proinflammatory cytokines , adhesion molecules , and matrix degrading enzymes . At the transcriptional level , they are regulated by the histone deacetylase sirtuin ( SIRT ) 1 via its actions on the proinflammatory transcription factor nuclear factor-κB ( NF-κB ) . The role of SIRT6 , also a histone deacetylase , in regulating inflammation in endothelial cells is not known . The aim of this study was to determine the effect of SIRT6 knockdown on inflammatory markers in human umbilical vein endothelial cells ( HUVECs ) in the presence of lipopolysaccharide ( LPS ) . LPS decreased expression of SIRT6 in HUVECs . Knockdown of SIRT6 increased the expression of proinflammatory cytokines ( IL-1β , IL-6 , IL-8 ) , COX-prostaglandin system , ECM remodelling enzymes ( MMP-2 , MMP-9 and PAI-1 ) , the adhesion molecule ICAM-1 , and proangiogenic growth factors VEGF and FGF-2 ; cell migration ; cell adhesion to leukocytes . Loss of SIRT6 increased the expression of NF-κB , whereas overexpression of SIRT6 was associated with decreased NF-κB transcriptional activity . Taken together , these results demonstrate that the loss of SIRT6 in endothelial cells is associated with upregulation of genes involved in inflammation , vascular remodelling , and angiogenesis . SIRT6 may be a potential pharmacological target for inflammatory vascular diseases . OUTPUT: activating invasion and metastasis;inducing angiogenesis;tumor promoting inflammation INPUT: Previous studies have demonstrated that the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone ( NNK ) induced liver tumors in F344 rats but not in Syrian golden hamsters . The aim of this study was to determine whether there was a correlation between the persistence of O6-methylguanine ( O6-mGua ) adducts and the rate of recovery of O6-methylguanine-DNA methyltransferase ( O6-mGuaT ) after depletion in the liver and susceptibility to NNK in F344 rat and Syrian golden hamster injected s.c. with NNK ( 80 mg/kg ) . The levels of both 7-methylguanine and O6-mGua reached a maximum 24 h after NNK treatment . O6-mGua in NNK-treated rat liver was undetectable after 48 h . In the rat , the depletion of O6-mGuaT activity occurred within 4 h following NNK treatment . A subsequent rapid recovery of enzyme activity was observed 36 h after NNK exposure . In contrast , high levels of O6-mGua persisted in hamster liver DNA and no O6-mGuaT activity was detected up to 336 h after NNK injection . Thus , the persistence of O6-mGua in hamster liver is most likely related to a lack of recovery of the O6-mGuaT . These results suggested that factors other than O6-mGua may be determining NNK-induced hepatocarcinogenesis in rats . An aldehyde generated by alpha-hydroxylation of NNK , 4-oxo-4-(3-pyridyl)butanal , inhibited O6-mGuaT activity in rat hepatocytes , suggesting that this aldehyde contributes to the carcinogenicity of NNK by inhibiting this repair enzyme . OUTPUT: genomic instability and mutation INPUT: It is prevailingly thought that the antiestrogens tamoxifen and ICI 182 , 780 are competitive antagonists of the estrogen-binding site of the estrogen receptor-alpha ( ER-α ) . However , a plethora of evidence demonstrated both antiestrogens exhibit agonist activities in different systems such as activation of the membrane-initiated signaling pathways . The mechanisms by which antiestrogens mediate estrogen-like activities have not been fully established . Previously , a variant of ER-α , EP-α36 , has been cloned and showed to mediate membrane-initiated estrogen and antiestrogen signaling in cells only expressing ER-α36 . Here , we investigated the molecular mechanisms underlying the antiestrogen signaling in ER-negative breast cancer MDA-MB-231 and MDA-MB-436 cells that express high levels of endogenous ER-α36 . We found that the effects of both 4-hydoxytamoxifen ( 4-OHT ) and ICI 182 , 780 ( ICI ) exhibited a non-monotonic , or biphasic dose response curve ; antiestrogens at low concentrations , elicited a mitogenic signaling pathway to stimulate cell proliferation while at high concentrations , antiestrogens inhibited cell growth . Antiestrogens at l nM induced the phosphorylation of the Src-Y416 residue , an event to activate Src , while at 5 �M induced Src-Y527 phosphorylation that inactivates Src . Antiestrogens at 1 nM also induced phosphorylation of the MAPK/ERK and activated the Cyclin D1 promoter activity through the Src/EGFR/STAT5 pathways but not at 5 �M . Knock-down of ER-α36 abrogated the biphasic antiestrogen signaling in these cells . Our results thus indicated that ER-α36 mediates biphasic antiestrogen signaling in the ER-negative breast cancer cells and Src functions as a switch of antiestrogen signaling dependent on concentrations of antiestrogens through the EGFR/STAT5 pathway . OUTPUT: sustaining proliferative signaling INPUT: AIM To investigate whether luteolin , a highly prevalent flavonoid , reverses the effects of epithelial-mesenchymal transition ( EMT ) in vitro and in vivo and to determine the mechanisms underlying this reversal . METHODS Murine malignant melanoma B16F10 cells were exposed to 1% O(2) for 24 h . Cellular mobility and adhesion were assessed using Boyden chamber transwell assay and cell adhesion assay , respectively . EMT-related proteins , such as E-cadherin and N-cadherin , were examined using Western blotting . Female C57BL/6 mice ( 6 to 8 weeks old ) were injected with B16F10 cells ( 1�10(6) cells in 0.2 mL per mouse ) via the lateral tail vein . The mice were treated with luteolin ( 10 or 20 mg/kg , ip ) daily for 23 d . On the 23rd day after tumor injection , the mice were sacrificed , and the lungs were collected , and metastatic foci in the lung surfaces were photographed . Tissue sections were analyzed with immunohistochemistry and HE staining . RESULTS Hypoxia changed the morphology of B16F10 cells in vitro from the cobblestone-like to mesenchymal-like strips , which was accompanied by increased cellular adhesion and invasion . Luteolin ( 5-50 μmol/L ) suppressed the hypoxia-induced changes in the cells in a dose-dependent manner . Hypoxia significantly decreased the expression of E-cadherin while increased the expression of N-cadherin in the cells ( indicating the occurrence of EMT-like transformation ) , which was reversed by luteolin ( 5 μmol/L ) . In B16F10 cells , luteolin up-regulated E-cadherin at least partly via inhibiting the β3 integrin/FAK signal pathway . In experimental metastasis model mice , treatment with luteolin ( 10 or 20 mg/kg ) reduced metastatic colonization in the lungs by 50% . Furthermore , the treatment increased the expression of E-cadherin while reduced the expression of vimentin and β3 integrin in the tumor tissues . CONCLUSION Luteolin inhibits the hypoxia-induced EMT in malignant melanoma cells both in vitro and in vivo via the regulation of β3 integrin , suggesting that luteolin may be applied as a potential anticancer chemopreventative and chemotherapeutic agent . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND Although chemotherapy offers promise of increased survival for children with medulloblastoma and glioblastoma multiforme , drug resistance occurs frequently , resulting in tumor progression and death . Resistance to nitrosoureas and methylating agents , which damage DNA , can be mediated by a DNA repair protein , O6-alkylguanine-DNA alkyltransferase ( AGAT ) . Depletion of this protein with alkylguanines or methylating agents , however , restores tumor cell sensitivity to the cytotoxicity of chloroethylnitrosoureas ( e.g. , carmustine [ BCNU] ) . PURPOSE This study was designed to determine whether resistance to the activity of nitrosourea ( the drug BCNU ) in BCNU-resistant human medulloblastoma ( D341 Med ) and human glioblastoma multiforme ( D-456 MG ) can be reversed by the methylating agent streptozocin and the O6-substituted guanines O6-methylguanine and O6-benzylguanine . METHODS Xenografts were grown subcutaneously in athymic BALB/c mice . BCNU was administered as a single intraperitoneal injection at doses of 100 mg/m2 , 75 mg/m2 , or 38 mg/m2--i.e. , 1.0 , 0.75 , or 0.38 , respectively , of the dose lethal to 10% of treated animals ( LD10 ) . Mice were treated intraperitoneally with a single dose of O6-benzylguanine or O6-methylguanine ( 240 mg/m2 ) or with streptozocin ( 600 mg/m2 ) daily for 4 days . Response was assessed by tumor growth delay and tumor regression . AGAT activity in the xenografts was measured at 1 and 6 hours after pretreatment , at the time tumors were excised . RESULTS Pretreatment with O6-benzylguanine , O6-methylguanine , or streptozocin reduced AGAT activity to 4% , 25% , and 95% of control values , respectively , in D341 Med and 0% , 0% , and 25% of control values , respectively , in D-456 MG 1 hour after injection . After 6 hours , levels changed to 7% , 61% , and 116% of control values in D341 Med and 0% , 79% , and 21% of control values in D-456 MG , respectively . Both D341 Med and D-456 MG xenografts were completely resistant to BCNU at its LD10 . Pretreatment with O6-benzylguanine increased BCNU sensitivity in both types of xenograft . In contrast , treatment with BCNU plus O6-methylguanine or streptozocin did not produce growth delays substantially different from those produced by BCNU alone , reflecting the more efficient depletion of AGAT by O6-benzylguanine . Following therapy with BCNU plus O6-benzylguanine at 0.38 LD10 , tumor regressions were seen in eight of 10 D341 Med and in all 10 D-456 MG xenografts . CONCLUSION We recommend comprehensive clinical toxicologic evaluation of combination therapy with O6-benzylguanine plus BCNU , which would allow subsequent design of phase I clinical trials . OUTPUT: genomic instability and mutation INPUT: A6 is an eight amino acid peptide derived from the non-receptor binding region of urokinase plasminogen activator ( uPA ) , which interferes with the uPA/uPA receptor system . A6 has been synthesized as a potential anti-angiogenic , anti-cancer agent . The current study has investigated the potential therapeutic activity of A6 in the Lewis lung carcinoma ( 3LL ) model of pulmonary metastasis . A6 was found to have direct anti-tumor activity against established 3LL pulmonary metastases at a low tumor burden ( 10-20 colonies per lung ) and was therapeutic in combination with cyclophosphamide at high tumor burdens ( &gt ; 100 colonies per lung ) . Mechanistic studies have revealed that A6 directly inhibits the invasion of 3LL cells through a Matrigel model basement membrane by 40-45% . Moreover , treatment with either A6 or doxorubicin resulted in thicker tubes in endothelial tube formation studies . Our results suggest that A6 , by virtue of its anti-invasive and anti-angiogenic properties , might work additively or synergistically with chemotherapeutic agents and thereby contribute to enhanced therapy of established 3LL cancer metastases . OUTPUT:

activating invasion and metastasis;inducing angiogenesis

HoC_dynamic_5_shot32

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Angiogenic factors produced by monocytes-macrophages are involved in the pathogenesis of chronic inflammatory disorders characterized by persistent angiogenesis . The possibility was tested that interleukin-8 ( IL-8 ) , which is a cytokine that is chemotactic for lymphocytes and neutrophils , is also angiogenic . Human recombinant IL-8 was potently angiogenic when implanted in the rat cornea and induced proliferation and chemotaxis of human umbilical vein endothelial cells . Angiogenic activity present in the conditioned media of inflamed human rheumatoid synovial tissue macrophages or lipopolysaccharide-stimulated blood monocytes was equally blocked by antibodies to either IL-8 or tumor necrosis factor-alpha . An IL-8 antisense oligonucleotide specifically blocked the production of monocyte-induced angiogenic activity . These data suggest a function for macrophage-derived IL-8 in angiogenesis-dependent disorders such as rheumatoid arthritis , tumor growth , and wound repair . OUTPUT: inducing angiogenesis INPUT: Although tumor-associated macrophages ( TAMs ) are involved in tumor growth and metastasis , the mechanisms controlling their pro-tumoral activities remain largely unknown . The transcription factor c-MYC has been recently shown to regulate in vitro human macrophage polarization and be expressed in macrophages infiltrating human tumors . In this study , we exploited the predominant expression of LysM in myeloid cells to generate c-Myc(fl/fl) LysM(cre/+) mice , which lack c-Myc in macrophages , to investigate the role of macrophage c-MYC expression in cancer . Under steady-state conditions , immune system parameters in c-Myc(fl/fl) LysM(cre/+) mice appeared normal , including the abundance of different subsets of bone marrow hematopoietic stem cells , precursors and circulating cells , macrophage density , and immune organ structure . In a model of melanoma , however , TAMs lacking c-Myc displayed a delay in maturation and showed an attenuation of pro-tumoral functions ( e.g. , reduced expression of VEGF , MMP9 , and HIF1α ) that was associated with impaired tissue remodeling and angiogenesis and limited tumor growth in c-Myc(fl/fl) LysM(cre/+) mice . Macrophage c-Myc deletion also diminished fibrosarcoma growth . These data identify c-Myc as a positive regulator of the pro-tumoral program of TAMs and suggest c-Myc inactivation as an attractive target for anti-cancer therapy . OUTPUT: avoiding immune destruction INPUT: The involvement of PRL in regulating monocyte/macrophage functions is suggested by the presence of PRL-Rs in these cells . Here , we show that PRL , though it failed to activate mouse peritoneal resident macrophages ( RMs ) , acted as a second signal and activated mouse peritoneal inflammatory macrophages ( EMs ) to a tumoricidal state . The cytotoxicity of mouse tumor-associated macrophages ( TAMs ) isolated at day 1 of tumor ( Ehrlich ascites carcinoma , EAC ) growth was enhanced by PRL . However , with progression of tumor growth , TAMs became nonresponsive to the hormone . PRL-induced killing of P815 target cells by EMs and TAMs was independent of TNF but correlated with the hormone-induced augmentation of NO2(-) and O2(-) release in these macrophages . Administration of PRL in vivo inhibited EAC growth and augmented NO2(-) release by TAMs . PRL synergized with the TH1 cytokine IFN-gamma , a known activator of macrophages , in inducing tumor killing and release of NO2(-) from EMs and TAMs . The hormone might activate macrophages at least partially , through the release of IFN-gamma as anti-IFN-gamma blocked IFN-gamma- as well as PRL-induced cytotoxicity in EMs . The TH2 cytokine IL-4 suppressed PRL-induced activation of macrophages . PRL induced release of IL-12 from EMs also , which suggested that the hormone might drive the TH1 response through IL-12 . Our observations further suggest that PRL alone and in synergy with IFN-gamma , released through induction of IL-12 , may generate tumoricidal macrophages and thus regulate the antitumor immune response of tumor hosts . OUTPUT: tumor promoting inflammation INPUT: Tumor-infiltrating immune cells can promote chemoresistance and metastatic spread in aggressive tumors . Consequently , the type and quality of immune responses present in the neoplastic stroma are highly predictive of patient outcome in several cancer types . In addition to host immune responses , intrinsic tumor cell activities that mimic stem cell properties have been linked to chemoresistance , metastatic dissemination and the induction of immune suppression . Cancer stem cells are far from a static cell population ; rather , their presence appears to be controlled by highly dynamic processes that are dependent on cues from the tumor stroma . However , the impact immune responses have on tumor stem cell differentiation or expansion is not well understood . In this study , we demonstrate that targeting tumor-infiltrating macrophages and inflammatory monocytes by inhibiting either the myeloid cell receptors CSF1R or CCR2 decreases the number of tumor-initiating cells in pancreatic tumors . Targeting CCR2 or CSF1R improves chemotherapeutic efficacy , inhibits metastasis and increases antitumor T-cell responses . Tumor-educated macrophages also directly enhanced the tumor-initiating capacity of pancreatic tumor cells by activating the transcription factor STAT3 , thereby facilitating macrophage-mediated suppression of CD8+ T lymphocytes . Together , our findings show how targeting tumor-infiltrating macrophages can effectively overcome therapeutic resistance mediated by tumor-initiating cells . OUTPUT: tumor promoting inflammation;activating invasion and metastasis;avoiding immune destruction INPUT: Biallelic mutations in MCPH1 cause primary microcephaly ( MCPH ) with the cellular phenotype of defective chromosome condensation . MCPH1 encodes a multifunctional protein that notably is involved in brain development , regulation of chromosome condensation , and DNA damage response . In the present studies , we detected that MCPH1 encodes several distinct transcripts , including two major forms : full-length MCPH1 ( MCPH1-FL ) and a second transcript lacking the six 3 ' exons ( MCPH1Δe9-14 ) . Both variants show comparable tissue-specific expression patterns , demonstrate nuclear localization that is mediated independently via separate NLS motifs , and are more abundant in certain fetal than adult organs . In addition , the expression of either isoform complements the chromosome condensation defect found in genetically MCPH1-deficient or MCPH1 siRNA-depleted cells , demonstrating a redundancy of both MCPH1 isoforms for the regulation of chromosome condensation . Strikingly however , both transcripts are regulated antagonistically during cell-cycle progression and there are functional differences between the isoforms with regard to the DNA damage response ; MCPH1-FL localizes to phosphorylated H2AX repair foci following ionizing irradiation , while MCPH1Δe9-14 was evenly distributed in the nucleus . In summary , our results demonstrate here that MCPH1 encodes different isoforms that are differentially regulated at the transcript level and have different functions at the protein level . OUTPUT: genomic instability and mutation;sustaining proliferative signaling INPUT: Tumor-associated macrophages ( TAM ) have been shown to play an important role in tumor angiogenesis . The purpose of this study was to determine whether monocyte recruitment , activation and differentiation mediated by monocyte chemotactic protein-1 ( MCP-1 ) and macrophage colony stimulating factor ( M-CSF ) modulate the expression of the angiogenic factor , Interleukin ( IL)-8 . Isolated human peripheral blood monocytes secreted low basal levels of IL-8 . Incubation of monocytes with M-CSF or MCP-1 resulted in an up-regulation of IL-8 mRNA and protein expression . The differential expression of IL-8 by monocytes following MCP-1 and M-CSF treatments involved activation of the NFkB transcription factor . Further activation with lipopolysaccharide ( LPS ) caused an increase in IL-8 secretion in monocytes but not in monocyte-derived macrophages ( MDM ) . MDM-conditioned media significantly up-regulated IL-8 expression in human malignant melanoma cells in vitro . In summary , we demonstrated that MCP-1 and M-CSF , critical for monocyte recruitment , activation and differentiation , differentially regulate IL-8 expression and may play an important role in monocyte/macrophage-mediated tumor angiogenesis . OUTPUT:

inducing angiogenesis

HoC_dynamic_5_shot33

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Most ovarian cancers are estrogen-positive and hormonal treatments using anti-estrogens or aromatase inhibitors are under investigation for treating the tumors that are resistant to conventional therapies . In this study , the long-term effects of two anti-estrogens , namely 4-hydroxytamoxifen and fulvestrant ( or ICI182,780 ) , were investigated in ERα-positive BG1 epithelial ovarian cancer cells . To this aim , cells were grown in the presence of anti-estrogen concentrations that were sufficient to saturate the estrogen receptors , but were neither cytotoxic nor cytostatic as indicated by the absence of inhibition of cell proliferation . In these conditions and despite the lack of cytostatic effect of the drugs , long-term treatment ( 3 months ) with the pure anti-estrogen fulvestrant induced a specific , reproducible and irreversible inhibition of ERα expression . This inhibition was accompanied by loss of estrogen-induced cell proliferation and gene expression as indicated by the analysis of several estrogen-responsive genes . ERα down-regulation was not linked to deregulated expression of transcription factors which drive ERα transcription and did not involve DNA methylation or histone deacetylation . Altogether , these results demonstrate that non-cytotoxic concentrations of pure anti-estrogens affect estrogen signaling and might be relevant for the treatment for ovarian cancers . OUTPUT: sustaining proliferative signaling INPUT: INTRODUCTION HER2 and estrogen receptor ( ER ) are important in breast cancer and are therapeutic targets of trastuzumab ( Herceptin ) and tamoxifen , respectively . Retinoids inhibit breast cancer growth , and modulate signaling by HER2 and ER . We hypothesized that treatment with retinoids and simultaneous targeting of HER2 and/or ER may have enhanced anti-tumor effects . METHODS The effects of retinoids combined with trastuzumab or tamoxifen were examined in two human breast cancer cell lines in culture , BT474 and SKBR3 . Assays of proliferation , apoptosis , differentiation , cell cycle distribution , and receptor signaling were performed . RESULTS In HER2-overexpressing/ER-positive BT474 cells , combining all-trans retinoic acid ( atRA ) with tamoxifen or trastuzumab synergistically inhibited cell growth , and altered cell differentiation and cell cycle . Only atRA/trastuzumab-containing combinations induced apoptosis . BT474 and HER2-overexpressing/ER-negative SKBR3 cells were treated with a panel of retinoids ( atRA , 9-cis-retinoic acid , 13-cis-retinoic acid , or N-(4-hydroxyphenyl) retinamide ( fenretinide ) ( 4-HPR) ) combined with trastuzumab . In BT474 cells , none of the single agents except 4-HPR induced apoptosis , but again combinations of each retinoid with trastuzumab did induce apoptosis . In contrast , the single retinoid agents did cause apoptosis in SKBR3 cells ; this was only modestly enhanced by addition of trastuzumab . The retinoid drug combinations altered signaling by HER2 and ER . Retinoids were inactive in trastuzumab-resistant BT474 cells . CONCLUSIONS Combining retinoids with trastuzumab maximally inhibits cell growth and induces apoptosis in trastuzumab-sensitive cells . Treatment with such combinations may have benefit for breast cancer patients . OUTPUT: sustaining proliferative signaling;resisting cell death INPUT: Several studies have indicated that the cell-surface expressed nucleolin is implicated in tumorigenesis and angiogenesis , and represents an important target for cancer therapy . Here we show that treatment of rhabdoid tumor derived G401 cells with a nucleolin antagonist , the HB-19 pseudopeptide , could restore contact inhibition , impair anchorage-independent growth , and suppress tumor development in nude mice . G401 cells grow without contact inhibition , which is an in vitro characteristic property of malignant tumor cells . At concentrations of HB-19 that does not affect cell viability and multiplication index , there is restoration of contact inhibition thus suggesting that HB-19 treatment causes reversion of the malignant phenotype . Accordingly , HB-19 pretreated G401 cells lose the capacity to form colonies in soft agar . When assayed for tumorigenicity in nude mice , only 50% of mice injected with HB-19 pretreated G401 cells developed tumors with the mean tumor weight of 0.32 g , compared to 100% of mice injected with control G401 cells with the mean tumor weight of 2.36 g . Interestingly , the restoration of contact inhibition in HB-19 treated G401 cells is concomitant with marked reduction of transcripts coding the Wilms ' tumor 1 gene , matrix metalloproteinase-2 , epithelial isoform of CD44 , and vascular endothelial growth factor , whereas no apparent modification is detected for transcripts coding the proto-oncogene c-Myc , anti-apoptotic Bcl-2 , pro-apoptotic Bax , tissue inhibitor of metalloproteinase TIMP-1 , angiogenesis inhibitor TSP-1 , and growth factor Midkine . These findings indicate that the molecular mechanism of action of HB-19 on such highly malignant rhabdoid tumor cells is associated with a selective inhibitory effect on the expression of genes implicated in tumorigenesis and angiogenesis . OUTPUT: evading growth suppressors INPUT: Radix of Asiasarum heterotropoides var. mandshuricum F. Maekawa ( A. radix ) has been prescribed for treating pain , allergies and inflammatory disorders in traditional oriental medicine . However , only limited information on the anticancer effects of A. radix is currently available . The aim of this study was to determine the anticancer effect of the ethanol extract of A. radix ( EEAR ) on HCT-116 human colon cancer cells and to investigate its underlying mechanisms of action . EEAR significantly induced G2/M cell cycle arrest and apoptosis in HCT-116 cells . EEAR-induced apoptosis was observed in parallel with activation of caspases and an increased ratio of Bax ( pro-apoptotic)/Bcl2 ( anti-apoptotic ) . Western blot analyses revealed that EEAR elevated the expression of p53 and p21(Waf/Cip1) and decreased the expression of the regulator proteins of G2/M phase progression , such as cdc2 and cyclin B. The upregulation of p53 by EEAR was due to the increased levels of p53 mRNA without a similar increase in proteasome-mediated p53 degradation . EEAR-induced apoptosis in HCT-116 cells was dependent on p53 expression , as determined by siRNA-mediated p53 knockdown . Taken together , these results suggest that EEAR inhibits the growth of the HCT-116 cells through induction of G2/M cell cycle arrest and apoptosis , which are mediated by p53 expression . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: PURPOSE Tamoxifen , a selective oestrogen receptor modulator ( SERM ) , and brivanib alaninate , a vascular endothelial growth factor receptor 2 ( VEGFR-2 ) inhibitor , are two target specific agents that result in a substantial decrease in tumour growth when given alone . Tamoxifen activates SERM stimulated breast and endometrial tumour growth . Tamoxifen and brivanib alaninate have side-effects that can affect therapeutic outcomes . The primary goal of the current study was to evaluate the therapeutic effects of lower doses of both agents when given in combination to mice with SERM sensitive , oestrogen stimulated tumour xenografts ( MCF-7 E2 tumours ) . Experiments were conducted to evaluate the response of SERM stimulated breast ( MCF-7 Tam , MCF-7 Ral ) and endometrial tumours ( EnCa 101 ) to demonstrate the activity of brivanib alaninate in SERM resistant models . EXPERIMENTAL DESIGN In the current study , tumour xenografts were minced and bi-transplanted into the mammary fat pads of athymic , ovariectomised mice . Preliminary experiments were conducted to determine an effective oral dose of tamoxifen and brivanib alaninate that had minimal effect on tumour growth . Doses of 125 microg of tamoxifen and 0.05 mg/g of brivanib alaninate were evaluated . An experiment was designed to evaluate the effect of the two agents together when started at the time of tumour implantation . An additional experiment was done in which tumours were already established and then treated , to obtain enough tumour tissue for molecular analysis . RESULTS Brivanib alaninate was effective at inhibiting tumour growth in SERM sensitive ( MCF-7 E2 ) and SERM stimulated ( EnCa 101 , MCF-7 Ral , MCF-7 Tam ) models . The effect of the low dose drug combination as an anti-tumour strategy for SERM sensitive ( MCF-7 E2 ) in early treatment was as effective as higher doses of either drug used alone . In established tumours , the combination is successful at decreasing tumour growth , while neither agent alone is effective . Molecular analysis revealed a decreased phosphorylation of VEGFR-2 in tumours that were treated with brivanib alaninate and an increase in VEGFA transcription to compensate for the blockade of VEGFR-2 by increasing the transcription of VEGFA . Tamoxifen increases the phosphorylation of VEGFR-2 and this effect is abrogated by brivanib alaninate . There was also increased necrosis in tumours treated with brivanib alaninate . CONCLUSION Historically , tamoxifen has a role in blocking angiogenesis as well as the blockade of the ER . Tamoxifen and a low dose of an angiogenesis inhibitor , brivanib alaninate , can potentially be combined not only to maximise therapeutic efficacy but also to retard SERM resistant tumour growth . OUTPUT: resisting cell death;inducing angiogenesis;sustaining proliferative signaling INPUT: Recent studies have shown that the antiestrogen tamoxifen ( TAM ) can be used in the treatment of malignant neoplasms other than breast cancer . In the present study , we investigated the expression of estrogen receptor ( ER ) in six malignant rhabdoid tumor ( MRT ) cell lines . Alterations in MRT cell growth in response to estrogen or antiestrogens ( 4-hydroxytamoxifen ( 4-OHT ) , TAM , and ICI 182 780 ) were also investigated . RT-PCR and western blotting showed that ER-alpha was expressed in three of the six MRT cell lines . While 17-beta-estradiol ( E2 ) did not significantly alter MRT cell line proliferation , the hydroxylated tamoxifen metabolite 4-OHT significantly inhibited the growth of all 6 MRT cell lines . However , the steroidal antiestrogen ICI 182 780 did not alter the proliferation of any of the MRT cell lines. 4-OHT induced apoptosis in both ER-alpha-negative and ER-alpha-positive MRT cell lines , as assessed by nuclear morphology and DNA fragmentation . Neither growth inhibition nor induction of apoptosis due to 4-OHT was blocked by the addition of excess E2 . Our data suggested that 4-OHT induced cytotoxic effects against MRT cells , and that these effects were independent of ER expression . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot34

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Context:The ubiquitin-proteasome system and macroautophagy are two major pathways for intracellular protein degradation . Emerging lines of evidence have shown that blockade of ubiquitin-proteasome system by proteasome inhibitors activates macroautophagy.Objective:The purpose of this study was to determine the involvement of autophagy essential gene Beclin 1 in cytotoxicity of thyroid cancer cells mediated by proteasome inhibitors.Design:Autophagy was measured by acidic-trophic dye staining and EGF-LC3 distribution using fluorescence microscopy , as well as LC3-II transition using Western blot . To ascertain the effect of Beclin 1 , cells were transfected with Beclin 1 plasmid or shRNA against Beclin 1 . Cell viability and apoptotic cells were measured using MTT assay and flow cytometry , respectively.Results:Proteasome inhibitors decreased Beclin 1 expression . In addition , treatment with PI3K inhibitors 3-MA or wortmannin , as well as knockdown of Beclin 1 expression , was unable to affect autophagic responses mediated by proteasome inhibitors . Overexpression of Beclin 1 enhanced proteasome inhibitor-mediated cytotoxicity of thyroid cancer cells via suppression of survivin.Conclusions:Proteasome inhibitors cause Beclin 1-independent macroautophagic responses of thyroid cancer cells in a Beclin 1-independent manner . Beclin 1 possesses autophagy-independent antitumoral effects upon exposure of thyroid cancer cells to proteasome inhibitors . OUTPUT: resisting cell death INPUT: ABSTRACT : BACKGROUND : The ubiquitin-proteasome system and macroautophagy ( hereafter referred to autophagy ) are two complementary pathways for protein degradation . Emerging evidence suggests that proteasome inhibition might be a promising approach for tumor therapy . Accumulating data suggest that autophagy is activated as a compensatory mechanism upon proteasome activity is impaired . METHOD : Autophagy activation was measured using acridine orange staining and LC3 transition . Cell viability and apoptosis were measured using MTT assay and flow cytometry , respectively . Beclin 1 expression vectors or shRNA against Beclin 1 ( shBeclin 1 ) were transfected to investigate the role of Beclin 1 in autophagy activation and cytotoxicity of ovarian cancer cells induced by proteasome inhibitors . RESULTS : Proteasome inhibitors suppressed proliferation and induced autophagy in ovarian cancer cells . Neither phosphoinositide 3-kinase ( PI3K ) inhibitors nor shRNA against Beclin 1 could abolish the formation of acidic vacuoles and the processing of LC3 induced by proteasome inhibitors . Moreover , Beclin 1 overexpression enhanced anti-proliferative effects of proteasome inhibitors in ovarian cancer cells . CONCLUSIONS : For the first time , the current study demonstrated that proteasome inhibitors induced PI3K and Beclin 1-independent autophagy in ovarian cancer cells . In addition , this study revealed autophagy-independent tumor suppressive effects of Beclin 1 in ovarian cancer cells . OUTPUT: resisting cell death INPUT: The relationship between thyroid hormone ( triiodothyronine , T(3) ) and breast cancer is unclear . We studied the effect of the c-erbA/TR alpha proto-oncogene encoding a functional T(3) receptor ( TR alpha 1 ) , of its ligand T(3) , and of its retroviral , mutated counterpart , the v-erbA oncogene , on the proliferation capacity of nontumorigenic mammary epithelial cells ( EpH4 ) . We found that EpH4 cells expressing ectopically TR ( EpH4 + TR alpha 1 ) or v-erbA ( EpH4 + v-erbA ) proliferated faster than parental EpH4 cells that contained low levels of endogenous TR . T(3) inhibited DNA synthesis and proliferation in EpH4 + TR alpha 1 cells but not EpH4 or EpH4 + v-erbA cells . The study of cell-cycle genes showed that T(3) decreased cyclin D1 RNA and protein levels in EpH4 + TR alpha 1 cells . In addition , T(3) downregulated the expression of T1 , a gene that is overexpressed in human breast adenocarcinomas and is induced by mitogens , serum , and several oncogenes and cytokines . Inhibition of the T1 gene by T(3) required both de novo mRNA and protein synthesis . Furthermore , T(3) abolished the induction of T1 by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate and inhibited the activity of an activation protein 1-dependent promoter ( -73-Col-CAT ) in EpH4 + TR alpha 1 cells , suggesting that interference with activation protein 1 transcription factor plays a part in the inhibition of the T1 gene . Our results showed that T(3) reduced the proliferation of mammary epithelial cells and inhibited the expression of cyclin D1 and T1 genes . OUTPUT: sustaining proliferative signaling INPUT: Bcl-2/E1B 19-kDa interacting protein 3 ( BNIP3 ) is a proapoptotic protein whose expression level is often low in colorectal cancer ( CRC ) cells due to the BNIP3 gene promoter DNA methylation by DNA methyltransferase ( DNMT ) . It is known that chemotherapy and radiotherapy suppress CRC through inducing tumor apoptosis . However , the molecular mechanisms underlying chemotherapy and radiotherapy-induced apoptosis of CRC cells are not well defined . In this study , we observed that the expression level of BNIP3 in colon cancer cells was significantly increased by treatment with therapeutic agents and radiation in vitro . The BNIP3 protein level in CRC tissues from patients who received preoperative concurrent chemotherapy was significantly higher than in those who received surgery alone . Furthermore , treatment with chemotherapeutic agents and radiation significantly decreased the DNMT1 expression level and enzymatic activity . Both expression level and activity of DNMT1 were inversely correlated with the expression level of BNIP3 in colon carcinoma cells after treatment with chemotherapeutic agents and radiation . Consistent with increased BNIP3 expression , chemotherapeutic agents and radiation induced colon carcinoma cell apoptosis in a dose-dependent manner . Based on these observations , we conclude that chemotherapy and radiotherapy inhibit DNMT1 expression to upregulate BNIP3 expression to promote CRC cell apoptosis . And , BNIP3 may play a role in the caspase-dependent apoptosis pathways , mainly during treatment with chemotherapy and radiotherapy . OUTPUT: resisting cell death INPUT: Cyclin E2 , but not cyclin E1 , is included in several gene signatures that predict disease progression in either tamoxifen-resistant or metastatic breast cancer . We therefore examined the role of cyclin E2 in antiestrogen resistance in vitro and its potential for therapeutic targeting through cyclin-dependent kinase ( CDK ) inhibition . High expression of CCNE2 , but not CCNE1 , was characteristic of the luminal B and HER2 subtypes of breast cancer and was strongly predictive of shorter distant metastasis-free survival following endocrine therapy . After antiestrogen treatment of MCF-7 breast cancer cells , cyclin E2 mRNA and protein were downregulated and cyclin E2-CDK2 activity decreased . However , this regulation was lost in tamoxifen-resistant ( MCF-7 TAMR ) cells , which overexpressed cyclin E2 . Expression of either cyclin E1 or E2 in T-47D breast cancer cells conferred acute antiestrogen resistance , suggesting that cyclin E overexpression contributes to the antiestrogen resistance of tamoxifen-resistant cells . Ectopic expression of cyclin E1 or E2 also reduced sensitivity to CDK4 , but not CDK2 , inhibition . Proliferation of tamoxifen-resistant cells was inhibited by RNAi-mediated knockdown of cyclin E1 , cyclin E2 , or CDK2 . Furthermore , CDK2 inhibition of E-cyclin overexpressing cells and tamoxifen-resistant cells restored sensitivity to tamoxifen or CDK4 inhibition . Cyclin E2 overexpression is therefore a potential mechanism of resistance to both endocrine therapy and CDK4 inhibition . CDK2 inhibitors hold promise as a component of combination therapies in endocrine-resistant disease as they effectively inhibit cyclin E1 and E2 overexpressing cells and enhance the efficacy of other therapeutics . OUTPUT: sustaining proliferative signaling INPUT: The thyroid hormone ( T3 ) blocks proliferation and induces differentiation of neuroblastoma N2a-beta cells that overexpress the beta 1 isoform of the T3 receptor . An element in the region responsible for premature termination of transcription mediates a rapid repression of c-myc gene expression by T3 . The hormone also causes a decrease of cyclin D1 gene transcription , and is able to antagonize the activation of the cyclin D1 promoter by Ras . In addition , a strong and sustained increase of the levels of the cyclin kinase inhibitor ( CKI ) p27(Kip1) are found in T3-treated cells . The increased levels of p27(Kip1) lead to a marked inhibition of the kinase activity of the cyclin-CDK2 complexes . As a consequence of these changes , retinoblastoma proteins are hypophosphorylated in T3-treated N2a-beta cells , and progression through the restriction point in the cell cycle is blocked . OUTPUT:

sustaining proliferative signaling;evading growth suppressors

HoC_dynamic_5_shot35

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cytokines are known to play an important role in host defense by regulating the function , growth , and differentiation of the cells of the immune system . We hypothesize that , in the tumor microenvironment , tumor cells and resident tissue cells ( e.g. , fibroblasts ) also produce cytokines that may regulate the local immune response to tumors . Initially , homogenates of eight head and neck squamous cell carcinomas ( HNSCC ) were assayed for the presence of interleukin-1 ( IL-1 ) , interleukin-4 ( IL-4 ) , interleukin-6 ( IL-6 ) , and granulocyte-macrophage colony-stimulating factor ( GM-CSF ) to establish the presence of these cytokines in the tumors in vivo . We detected IL-1 in all tumor homogenates and IL-4 , IL-6 , and GM-CSF in some homogenates . To assess the ability of HNSCC to produce these cytokines , supernatants of short-term primary cultures of HNSCC were assayed for the same cytokines . No IL-1 was detected , although baseline levels of IL-4 , IL-6 , and GM-CSF were present . However , the stimulation of primary tumor cultures with exogenous IL-1 induced or significantly enhanced production of IL-4 ( p &lt ; 0.01 ) , IL-6 ( p &lt ; 0.001 ) , and GM-CSF ( p &lt ; 0.02 ) . These results support our hypothesis that HNSCC secrete cytokines that may influence the response of local immune cells . Our data also suggest that IL-1 may have a central role in regulating the local immune response through the enhancement or induction of cytokine production by tumor and/or resident tissue cells . OUTPUT: avoiding immune destruction;tumor promoting inflammation INPUT: The CD4+CD25+ regulatory T cell ( Treg ) is a special kind of T cell subset . Studies have showed that Treg cells are involved in a number of physiological processes and pathologic conditions such as autoimmune diseases , transplantation tolerance and cancer . Tregs with unique capacity for immune inhibition can impair anti-tumour immunity and help tumor cells to escape from immune surveillance . The aim of our study was to investigate whether Tregs are involved in hepatocellular carcinoma ( HCC ) . A BABL/C mouse with HCC in situ model was established to evaluate the Treg existence in carcinoma tissues and the changes of Tregs in spleen using flow cytometry and immunohistochemistry methods . Granzyme B expression in carcinoma tissues was analyzed by immunohistochemistry to investigate the tumor local immune status . The proportion of CD4+CD25+/CD4+ spleen lymphocytes of tumor bearing mice ( 18.8% � 1.26% ) was found to be significantly higher than that in normal mice ( 9.99% � 1.90% ) ( P<0.01 ) . Immunohistochemistry of spleen tissue also confirmed that there was an increase in Treg in tumor-bearing mice , while in carcinomas it showed Treg cells to be present in tumor infiltrating lymphocyte areas while Granzyme B was rarely observed . Anti-tumour immunity was suppressed , and this might be associated with the increase of Tregs . Our observations suggest that the CD4+CD25+Treg/ CD4+ proportion in spleen lymphocytes can be a sensitive index to evaluate the change of Tregs in hepatocellular carcinoma mice and the Treg may be a promising therapeutic target for cancer . OUTPUT: avoiding immune destruction INPUT: Thyroid hormone ( T(3) ) mediates cellular growth , development , and differentiation by binding to the nuclear thyroid hormone receptor ( TR ) . Recent studies suggest that long-term hypothyroidism is associated with human hepatocellular carcinoma ( HCC ) independent from other major HCC risk factors . Dickkopf ( DKK ) 4 , a secreted protein , antagonizes the Wnt signal pathway . In this study , we demonstrate that T(3) may play a suppressor role by inducing DKK4 expression in HCC cells at both the messenger RNA ( mRNA ) and protein levels . DKK4 was down-regulated in 67.5% of HCC cancerous tissues . The decrease in DKK4 levels was accompanied by a concomitant decrease in TR protein levels in the matched cancerous tissues in 31% of tissues compared by immunoblotting with the adjacent noncancerous tissues . Further , TR and DKK4 expression levels were positively correlated in both normal and cancerous specimens by tissue array analysis . In function assays , stable DKK4 transfected into J7 or HepG2 cells decreased cell invasion in vitro . Conversely , knocking down DKK4 restores cell invasiveness . DKK4-expressing J7 clones showed increased degradation of β-catenin , but down-regulation of CD44 , cyclin D1 , and c-Jun . To investigate the effect of DKK4 and TR on tumor growth in vivo , we established a xenograft of J7 cells in nude mice . J7-DKK4 and J7-TRα1 overexpressing mice , which displayed growth arrest , lower lung colony formation index , and smaller tumor size than in control mice , supporting an inhibitory role of DKK4 in tumor progression . CONCLUSION : Taken together , these data suggest that the TR/DKK4/Wnt/β-catenin cascade influences the proliferation and migration of hepatoma cells during the metastasis process and support a tumor suppressor role of the TR . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND Hepatocellular carcinoma ( HCC ) is the most common liver cancer . Therapeutic results are usually unsatisfactory because liver tumors recur often . Immunologic factors may be related to the recurrence of HCC ; however , this possibility is mentioned only rarely . METHODS Thirty HCC patients undergoing hepatectomies were divided into 3 groups according to the diameters of their HCCs : group A ( n = 8 ) , diameter ≤3 cm ; group B ( n = 8 ) , diameter >3 cm and ≤5 cm ; and group C ( n = 14 ) , diameter >5 cm . T-lymphocytes from peripheral blood , nontumor liver tissue , and the HCC were analyzed . RESULTS The percentage of CD25+ in the CD4+ T cells did not differ between the peripheral blood and the nontumor liver tissue among the 3 groups . CD25+ cells were increased in the tumor tissue in group C patients ( range , 6-41% ; median , 22.9% ; P = .003 ) , compared to group A patients . The percentage of CD25+ in the CD4+ T cells in tumor tissue was positively correlated with tumor sizes ( r = 0.556 ) . These CD4+ CD25+ lymphocytes produced transforming growth factor-β and interferon-γ but not interleukin-10 , and were anergic to plate-coated monoclonal antibodies ( anti-CD3/anti-CD28 ) . The characteristics of these antibodies were comparable to those of regulatory T cells . When the infiltration lymphocytes including CD4+ CD25+ T cells were added to the mixed lymphocyte reaction activated by autologous tumor lysate-pulsed dendritic cells , the proliferation of lymphocytes was inhibited . CONCLUSION The increase of CD4+ CD25+ T cells in the tumor microenvironment correlates with tumor sizes . These CD4+ CD25+ regulatory T cells appeared to suppress the immune response activated by dendritic cells . OUTPUT: tumor promoting inflammation INPUT: While human embryonic stem cells ( hESCs ) and human embryonal carcinoma cells ( hECCs ) have been studied extensively at the levels of the genome , transcriptome , proteome and epigenome our knowledge of their corresponding metabolomes is limited . Here , we present the metabolic signatures of hESCs and hESCs obtained by untargeted gas chromatography coupled to mass spectrometry ( GC-MS ) . Whilst some metabolites are common to both cell types , representing the self-renewal and house-keeping signatures , others were either higher ( e.g. , octadecenoic acid , glycerol-3-phosphate , 4-hydroxyproline ) or lower ( e.g. , glutamic acid , mannitol , malic acid , GABA ) in hESCs ( H9 ) compared to hECCs ( NTERA2 ) , these represent cell type specific signatures . Further , our combined results of GC-MS and microarray based gene expression profiling of undifferentiated and OCT4-depleted hESCs are consistent with the Warburg effect which is increased glycolysis in embryonic cells and tumor cells in the presence of O(2) while oxidative phosphorylation ( OXPHOS ) is impaired or even shut down . RNAi-based OCT4 knock down mediated differentiation resulted in the activation of the poised OXPHOS machinery by expressing missing key proteins such as NDUFC1 , UQCRB and COX , increase in TCA cycle activity and decreased lactate metabolism . These results shed light on the metabolite layer of pluripotent stem cells and could potentially establish novel metabolic markers of self renewal and pluripotency . OUTPUT: cellular energetics INPUT: Little is known about the requirements for human T-cell leukemia virus type I ( HTLV-I ) entry , including the identity of the cellular receptor(s) . Recently , we have generated an HTLV-I surface glycoprotein ( SU ) immunoadhesin , HTSU-IgG , which binds specifically to cell-surface protein(s) critical for HTLV-I-mediated entry in cell lines . Here , expression of the HTLV-I SU binding protein on primary cells of the immune system was examined . The immunoadhesin specifically bound to adult T cells , B cells , NK cells , and macrophages . Cell stimulation dramatically increased the amount of binding , with the highest levels of binding on CD4(+) and CD8(+) T cells . Naive ( CD45RA(high) , CD62L(high) ) CD4(+) T cells derived from cord blood cells , in contrast to other primary cells and all cell lines examined , bound no detectable HTLV-I SU . However , following stimulation , the level of HTSU-IgG binding was rapidly induced ( fewer than 6 hours ) , reaching the level of binding seen on adult CD4(+) T cells by 72 hours . In contrast to HTLV-I virions , the soluble HTSU-IgG did not effect T-cell activation or proliferation . When incubated with human peripheral blood mononuclear cells in a mixed leukocyte reaction , HTSU-IgG inhibited proliferation at less than 1 ng/mL . These results indicate that cell-surface expression of the HTLV SU binding protein is up-regulated during in vitro activation and suggest a role for the HTLV-I SU binding proteins in the immunobiology of CD4(+) T cells . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot36

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: OBJECTIVES Lymph node metastasis is among the most important prognostic factors for patients with esophageal squamous cell carcinoma after curative esophagectomy ; however , the extent of lymphadenectomy is still controversial . The objective of the present study was to determine the frequency of lymphatic metastases and to study the pattern of lymph node metastasis in a large study population . METHODS The data from 1361 patients with thoracic esophageal squamous cell carcinoma who underwent curative R0 esophagectomy were retrospectively examined . Logistic regression analysis was used to identify the factors associated with lymph node metastasis . RESULTS Of the 1361 patients , 714 ( 52.5% ) were found to have lymph node metastasis . The frequency of lymph node metastasis increased as the tumor invasion increased . Paratracheal nodes were the most frequent metastasis nodes ( 15.9% ) . The frequency of lymph node metastasis was 9.8% in the neck , 18.0% in the upper mediastinum , 18.9% in the middle mediastinum , 11.8% in the lower mediastinum , and 28.4% in the abdomen . Of these 714 patients , 424 ( 31.2% ) presented with 1 field involvement , 255 ( 18.7% ) with 2 fields , and 35 ( 2.6% ) with 3 fields involvement . Logistic regression analysis revealed tumor length ( P<.001 ) , tumor invasion ( P<.001 ) , tumor differentiation ( P=.003 ) , and lymphovascular invasion ( P<.001 ) were risk factors for lymph node metastasis . Tumor location ( P<.001 ) , tumor invasion ( P=.003 ) , lymphovascular invasion ( P=.004 ) , and paratracheal lymph node involvement ( P=.002 ) were identified as risk factors for cervical lymph node metastasis . CONCLUSIONS Metastases were more frequent in the abdomen than in the neck . Total mediastinal and upper abdominal lymphadenectomy should be carefully conducted . Certain factors , such as tumor location , depth of tumor invasion , lymphovascular invasion , and paratracheal lymph node involvement , might be helpful in determining the need to perform cervical lymphadenectomy in individual patients . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND &amp ; OBJECTIVE Altered cell adhesion has a critical role in the development of epithelial cancers . E-cadherin act on the maintenance of cell-cell adhesion and its function was thought to be regulated by associated cytoplasmic proteins , such as alpha-catenin and beta-catenin . This study was designed to examine the expression of beta-catenin in gastric carcinoma and to determine the relationship between tumor characteristics and survival . METHODS Immunohistochemical staining of beta-catenin and E-cadherin was performed in 148 patients with gastric carcinoma . RESULTS Abnormal expression of beta-catenin and E-cadherin was performed in 148 patients with gastric carcinoma . RESULTS Abnormal expression of beta-catenin and E-cadherin was demonstrated in 43.2% and 44.6% of tumors respectively . Up to 63% of tumors stained abnormally for one or two components of the cadherin-catenin complex ( beta-catenin , E-cadherin ) . Abnormal beta-catenin and E-cadherin staining occurred more frequently in poor differentiated tumor than in good differentiated tumors ( P &lt ; 0.005 , respectively ) . There was a significant correlation between abnormal beta-catenin expression and depth of invasion(P &lt ; 0.025 ) . Moreover , abnormal beta-catenin expression was more frequent in tumors with positive lymph node metastasis ( 45/84 , 53.6% ) and distance metastasis(21/31 , 67.7% ) than in tumors without lymph node metastasis ( 19/64 , 29.7% ) ( P &lt ; 0.005 ) and distance metastasis ( 43/117 , 36.8% ) ( P &lt ; 0.005 ) . A survival advantage was noted in tumors retaining normal membranous expression of beta-catenin , independent of type , grade , or stage ( P &lt ; 0.005 ) . CONCLUSIONS Abnormal expression of the E-cadherin-catenin complex occurs frequently in gastric carcinoma . The close correlation with poor survival suggests that abnormal beta-catenin may be a useful prognostic marker . OUTPUT: activating invasion and metastasis INPUT: Metastasis is a major cause of death of patients with malignant tumors . Matrix metalloproteinases ( MMPs ) are important for the migration and invasion of various types of cancer cell . Propofol is a known anesthetic agent , widely used for short-term anesthesia and for longer-term sedation . Propofol inhibits the proliferation of a variety of tumor cells , but there is no available information regarding propofol-inhibited migration and invasion of tumor cells in vitro . In this study , we investigated the effects of propofol on the migration and invasion of human lung carcinoma A549 cells . Wound healing assay and Boyden chamber assays indicated that propofol inhibited the migration and invasion of A549 cells in vitro . Gelatin zymographic analysis showed the inhibitory effect of propofol on the activation of expression MMP-2 . Western blot analysis also indicated that propofol suppressed the protein expiration of growth factor receptor-bound protein 2 ( GRB2 ) , Jun N-terminal kinases 1/2 ( p-JNK1/2 ) , p-p38 , MMP-2 and MMP-9 in A549 cells . Results from real-time PCR assay also showed that propofol inhibited the mRNA gene expression of MMP-2 , -7 and -9 , and enhanced that of tissue inhibitor of metalloproteinase 1 ( TIMP1 ) and TIMP2 in A549 cells . Taken together , these data show that propofol inhibits MMP-2 and -9 mRNA and protein expressions , resulting in suppression of lung cancer cell invasion and migration in vitro . OUTPUT: activating invasion and metastasis INPUT: c-myc , c-erbB-2 , and Ki-67 expression was examined by immunohistochemistry in 11 normal breast tissues and 42 invasive and 14 noninvasive breast carcinomas . The c-myc product was detected in all breast carcinoma specimens and in 7 of 11 normal breast tissues . Invasive tumors stained more frequently with the anti-myc monoclonal antibody than did noninvasive tumors , while the level of expression in normal breast tissue was much less than that in breast cancer . Membrane staining of the c-erbB-2 protein was demonstrated in 29% ( 4 of 14 ) of noninvasive ductal carcinomas and in 45% ( 19 of 42 ) of invasive breast carcinomas . None of the 11 normal breast tissue samples was positive . The mean value of Ki-67-positive cells was 0.91 +/- 0.31% for normal breast tissue , 4.57 +/- 1.36% for noninvasive ductal carcinoma , and 12.76 +/- 2.18% for invasive breast cancer . In 42 invasive breast carcinomas , the expression of c-myc , c-erbB-2 , and Ki-67 proliferation marker were compared with lymph node status , estrogen receptor status , progesterone receptor status , and age of patients at diagnosis. c-erbB-2 overexpression and Ki-67 overexpression were identified as the only factors associated with lymph node status . We concluded that they might be additional prognostic factors for breast carcinoma . OUTPUT: sustaining proliferative signaling INPUT: BACKGROUND Adenoid cystic carcinoma ( ACC ) of the Breast is a rare tumour ( less than 1 % of all breast carcinomas ) . The aim of this study was to determine the clinical , histological and immunohistochemical characteristics of these tumours . METHODS From the database of the Bergonié Institute of Bordeaux , 30 cases of ACC were identified . The clinical and histological features of these carcinomas were characterized . An immunohistochemical study was performed with the following antibodies : ER , PR , HER-2-neu , Vimentin , EGFR , P63 , SMA , CK5/6 , CK8/18 , CK14 , cKIT , MIB1 , CD44 and CD24 . RESULTS Thirty patients were included ( median age 60.7 years ) . The 10 axillary lymph node dissections and two sentinel lymph procedures were negative . The architecture was frequently of a mixed type ( 26/30 ) and less often solid ( 4/30 ) . Among the 23 patients for whom follow up was available ( median follow-up : 84 months [ 2-288] ) , there were three local recurrences and three metastatic events . The tumors with recurrence and metastasis showed more necrosis , a mitotic count greater than 4/10hpf , and in one case perineural infiltration . All the tumours were ER , PR and Her-2-neu negative . Morphological and immunophenotypical analysis disclosed in each tumor , a basaloid and a luminal cell population with divergent immunophenotypical patterns . CONCLUSIONS The mammary ACC is made of two cell types and is of good prognosis despite its triple negative phenotype , similar to the basal-like infiltrating carcinoma NOS . Axillary lymph node dissection is not recommended . Good local control by at least large lumpectomy with long-term follow-up is necessary . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: BACKGROUND &amp ; OBJECTIVE There is little ideal predictor available on evaluating the lymph node metastatic potential of breast carcinoma . This study was designed to determine the expression of gene products of E-cadherin ( epithelial ) , N-cadherin ( nerve ) , and matrix metalloproteinase-9 ( MMP-9 ) in breast carcinoma tissue and investigate their association with the invasion and metastasis of breast carcinoma . METHODS The authors examined the expressions of E-cadherin , N-cadherin , and MMP-9 in 72 cases of breast carcinoma(39 cases with lymph node metastasis and 33 cases without lymph node metastasis ) by immunohistochemistry . Multivariable Cox proportional hazards model was used to analyze the patients ' prognosis . RESULTS The average ranks of E-cadherin in lymph node metastasis group and no lymph node metastasis group were 29.19 and 45.14 , respectively , with significant difference ( P &lt ; 0.001 ) . The expression of E-cadherin was correlated inversely with the metastasis of breast carcinoma . The average ranks of N-cadherin and MMP-9 were 40.04 and 42.97 in lymph node metastasis group , and 32.32 and 28.85 in no lymph node metastasis group , both with significant difference ( P &lt ; 0.05 ) , and these expressions were positively correlated with the lymph node metastasis of breast carcinoma . The patients who had high expression of E-cadherin had a longer survival time . CONCLUSION Expression of E-cadherin , N-cadherin , and MMP-9 are associated strongly with lymph node metastasis of breast carcinoma . These proteins are indicators of metastasis potential and prognosis of breast carcinoma . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot37

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Metastasis is a major cause of death of patients with malignant tumors . Matrix metalloproteinases ( MMPs ) are important for the migration and invasion of various types of cancer cell . Propofol is a known anesthetic agent , widely used for short-term anesthesia and for longer-term sedation . Propofol inhibits the proliferation of a variety of tumor cells , but there is no available information regarding propofol-inhibited migration and invasion of tumor cells in vitro . In this study , we investigated the effects of propofol on the migration and invasion of human lung carcinoma A549 cells . Wound healing assay and Boyden chamber assays indicated that propofol inhibited the migration and invasion of A549 cells in vitro . Gelatin zymographic analysis showed the inhibitory effect of propofol on the activation of expression MMP-2 . Western blot analysis also indicated that propofol suppressed the protein expiration of growth factor receptor-bound protein 2 ( GRB2 ) , Jun N-terminal kinases 1/2 ( p-JNK1/2 ) , p-p38 , MMP-2 and MMP-9 in A549 cells . Results from real-time PCR assay also showed that propofol inhibited the mRNA gene expression of MMP-2 , -7 and -9 , and enhanced that of tissue inhibitor of metalloproteinase 1 ( TIMP1 ) and TIMP2 in A549 cells . Taken together , these data show that propofol inhibits MMP-2 and -9 mRNA and protein expressions , resulting in suppression of lung cancer cell invasion and migration in vitro . OUTPUT: activating invasion and metastasis INPUT: OBJECTIVE To observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe ( RSR ) on HER2 positive breast cancer , to evaluate the effects of RSR on the expressions of matrix metalloproteinases ( MMPs ) and the tissue inhibitor of metalloproteinases ( TIMPs ) in the recurrence and metastasis of HER2 positive breast cancer , thus revealing its anti-tumor recurrent and metastatic mechanisms . METHODS Selected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu . The primary tumor resection was carried out . After surgery they were randomly divided into the blank control group , the RSR group , the Herceptin group , and the combination group ( RSR + Herceptin group ) . The treatment lasted for 4 months . The inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated . The expressions of matrix metalloproteinase-2 ( MMP-2 ) , matrix metalloproteinase-9 ( MMP-9 ) , tissue inhibitor of metalloproteinase ( TIMP-1 ) , and TIMP-2 in the recurrent tumor tissue were detected using Western blot . RESULTS By the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group , showing statistical difference between the two groups ( P = 0.037 ) . The average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group . The inhibition rate of lung metastasis was 37. 85% in the RSR group , but with no statistical significance . The expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group , the Herceptin group , and the combination group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group , the Herceptin group , and the combination group was significantly down-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group ( P &lt ; 0.05 ) . The expressions of both TIMP-1 and TIMP-2 of the RSR group , the Herceptin group , and the combination group were all up-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group ( P &lt ; 0.05 ) . It was higher in the combination group than in the RSR group ( P &lt ; 0.05 ) . CONCLUSIONS RSR could inhibit the tumor recurrence of FVB/neu mice . It could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix . It might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells . OUTPUT: activating invasion and metastasis INPUT: Background and Objective . The cell cycle is regulated by proteins at different checkpoints , and dysregulation of this cycle plays a role in carcinogenesis . Matrix metalloproteinases ( MMPs ) are enzymes that degrade collagen and promote tumour infiltration . The aim of this study was to evaluate the expression of various cell cycle regulators and MMPs and to correlate such expression with progression and recurrence in patients with stage T1 urothelial carcinoma of the bladder ( UCB ) . Patients and Methods . This population-based cohort study comprised 201 well-characterized patients with primary stage T1 urothelial carcinoma of the bladder . Immunohistochemistry was performed on formalin-fixed material to quantify expression of cell cycle regulators and two MMPs . Results . Normal expression of p53 and abnormal expression of MMP9 were associated with greater risk of tumour recurrence . Also , normal p16 expression was related to a lower risk of tumour progression . MMP2 , p21 , cyclin D1 , and pRb showed no significant results that could estimate progression or recurrence . Conclusions . Normal p16 expression is associated with a lower risk of tumour progression , but immunohistochemistry on cell cycle regulators and MMPs has little value in predicting the prognosis in stage T1 UCB . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Adipose tissue growth and development are thought to be associated with angiogenesis and extracellular matrix remodeling . Because ginseng has been shown to inhibit angiogenesis and matrix metalloproteinase ( MMP ) activity , we hypothesized that adipose tissue growth and obesity can be regulated by Korean ginseng ( Panax ginseng C.A. Meyer ) . Wild-type C57BL/6J mice were fed for 8 weeks with a low fat diet , a high fat diet ( HFD ) , or HFD supplemented with 0.5% or 5% Korean red ginseng extract . We measured body weight , adipose tissue mass , food intake , MMP activity , and the expression of genes involved in angiogenesis and MMPs . Administering ginseng to HFD-induced obese mice produced reductions in body weight and adipose tissue mass compared with untreated counterparts . Ginseng treatment decreased blood vessel density and MMP activity in adipose tissues . Ginseng also reduced mRNA levels of angiogenic factors ( e.g. , VEGF-A and FGF-2 ) and MMPs ( e.g. , MMP-2 and MMP-9 ) , whereas it increased mRNA levels of angiogenic inhibitors ( e.g. , TSP-1 , TIMP-1 , and TIMP-2 ) in adipose tissues . These results demonstrate that ginseng effectively reduces adipose tissue mass and prevents obesity in diet-induced obese mice and that this process may be mediated in part through the anti-angiogenic actions of ginseng . OUTPUT: inducing angiogenesis INPUT: BACKGROUND Peritoneal transport status is important not only for prescription , but also as a prognostic index . Flt-1 and Flk-1 , the major vascular endothelial growth factor receptors involved in angiogenesis and hyperpermeability , may play a potent role in determining peritoneal transport characteristics . However , the relationship between them has not been studied to date . We hypothesized that Flt-1 and Flk-1 expression in the peritoneal vasculature of uremic patients could be closely related to baseline peritoneal transport status . METHODS Thirty-six new patients without a previous history of peritonitis were enrolled . Clinical parameters such as age , sex , height , weight , causes of renal failure , and residual renal function were assessed . Parietal peritoneal biopsies were obtained during implantation of peritoneal dialytic catheters . Flt-1 and Flk-1 were semi-quantitatively evaluated by immunohistochemical staining . Peritoneal microvascular density ( MVD ) was counted . Within 6 weeks after commencing peritoneal dialysis , a standard peritoneal equilibration test was performed , and the dialysate-to-plasma concentration ratio for creatinine at 4 h ( D4/P Cr ) was determined . The patients were divided into two groups based on the D4/P Cr : more than 0.65 ( Group H , n = 22 ) and less than or equal to 0.65 ( Group L , n = 14 ) . The 24-h peritoneal protein excretion ( PPE ) was assayed . Flt-1 and Flk-1 were correlated with peritoneal MVD , D4/P Cr , and PPE . RESULTS Flt-1 and Flk-1 were detected in the peritoneal vasculature of uremic patients . Flt-1 expression was similar between the two groups , but Flk-1 expression in Group H was significantly higher than that in Group L ( p = 0.001 ) . Flt-1 expression did not show significant correlations with peritoneal MVD , D4/P Cr , and PPE . However , Flk-1 expression showed significant correlations with the above three parameters ( p &lt ; 0.001 for all ) . CONCLUSIONS For the first time , the expressions of Flt-1 and Flk-1 in peritoneal vasculature of uremic patients were detected . Flk-1 expression in peritoneal vasculature of uremic patients is closely correlated with the number of peritoneal microvessels , peritoneal small solute transport rate , and PPE . Our findings strongly suggest that Flk-1 may be a crucial determinant of baseline peritoneal transport characteristics . Further interventional studies are needed . OUTPUT: inducing angiogenesis INPUT: BACKGROUND &amp ; OBJECTIVE Usually pituitary adenomas are histological benign and grow slowly , but a proportion of them will become locally aggressive , and develop into invasive pituitary adenomas . The reasons for these differences in tumor behavior are poorly understood . Pituitary adenomas are abounding blood vessels . Angiogenesis and tumor invasion both require degradation of the extracellular matrix components to allow cell migration . The matrix metalloproteinases ( MMPs ) and their nature inhibitors-the tissue inhibitors of metalloproteinases ( TIMPs ) may play a central role in these processes . The aggressive mechanism of pituitary adenomas was studied through investigating the expression of MMP-9 , MMP-2 , TIMP-1 , and TIMP-2 in both invasive and non-invasive adenomas . METHODS Sixty-one surgical removed pituitary adenomas ( forty-nine cases invasive and twelve non-invasive adenomas ) were investigated . Immunohistochemistry staining ( SP method ) was used to detect the expression of MMP-9 , MMP-2 , TIMP-1 , and TIMP-2 in two groups . The results were treated with semi-quantitative method and analyzed by using non-parameter rank sum test . RESULTS Immunohistochemical staining of tumor cells for MMP-9 , TIMP-1 , MMP-2 , and TIMP-2 were noted 95.9% ( 47/49 ) , 57.1% ( 28/49 ) , 75.5% ( 37/49 ) and 89.8% ( 44/49 ) in invasive adenomas , and 100% ( 12/12 ) , 91.7% ( 11/12 ) , 66.7% ( 8/12 ) , and 91.7% ( 11/12 ) in non-invasive adenomas , respectively . Invasive tumors were significantly less expressing TIMP-1 and TIMP-2 ( P &lt ; 0.05 ) . There was no significant difference for MMP-9 or MMP-2 between invasive and non-invasive groups ( P &gt ; 0.05 ) . CONCLUSIONS TIMP-1 and TIMP-2 may play a key role in invasive pituitary adenomas to biological behavior . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot38

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND : CD81 is a transmembrane protein that serves as a putative receptor for hepatitis C virus . In addition , CD81 has been suggested to be involved in a broad range of other cellular functions . Its putative implication in tumorigenesis has so far , however , remained largely unexplored . To assess the candidacy of CD81 as a tumor suppressor in gastric cancer development , we investigated its expression and function in a series of primary gastric tumors and gastric tumor-derived cell lines . METHODS : The expression and concomitant methylation status of the CD81 gene and its effect on tumor development and cellular signaling were evaluated . RESULTS : CD81 mRNA levels were found to be low in 16 of 40 ( 40% ) primary tumors and 9 of 14 ( 64.2% ) cell lines , and these low expression levels were found to correlate with the stage and grade of the tumors . Genomic alterations of CD81 were not encountered , whereas its expression could be re-activated in low expressing cells upon 5-aza-dC treatment . Bisulfite DNA sequencing analysis of 10 CpG sites within the 5 ' proximal region of the CD81 gene promoter revealed that the observed transcriptional silencing was tightly associated with aberrant hypermethylation . Subsequent restoration of CD81 expression induced a G(1) cell cycle arrest and apoptosis , whereas siRNA-mediated CD81 down-regulation promoted cell proliferation and attenuated cellular responses to various apoptotic stress stimuli . Also the colony-forming ability of the tumor cells could be inhibited and enhanced through CD81 up- and down-regulation , respectively . CD81 was found to inhibit p38 ( but not ERK , JNK and AKT ) phosphorylation and its growth suppressive effect could be abolished through p38 up- and down-regulation . CONCLUSION : From our data we conclude that epigenetic inactivation of CD81 is a common feature of gastric tumors and that this inactivation may render growth and survival advantages to the tumor cells , at least partially through p38 signaling . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: Geraniol ( GOH ) , a naturally occurring monoterpene , has been shown to have antiproliferative , cell cycle arrest and apoptosis-inducing effects , and represents a promising cancer chemopreventive agent . In the present study , we investigated the chemopreventive potential of GOH ( 50 and 100 mg kg(-1) body weight ) against 7,12-dimethylbenz[a]anthracene ( DMBA)/12-O-tetradecanoylphorbol 13-acetate ( TPA)-mediated skin tumorigenesis in Swiss albino mice . The topical treatment of GOH , 30 min prior to TPA ( 2 µg per 200 µl of acetone ) treatment significantly inhibited TPA-induced skin edema , hyperplasia , COX-2 induction and oxidative stress response . The GOH treatment also resulted in reduction of TPA-induced ornithine decarboxylase activity and [ (3) H ] thymidine incorporation by 53% ( P < 0.001 ) and 41% ( P < 0.001 ) , respectively . We found that GOH treatment significantly inhibited the tumor incidence and number of tumors ( P < 0.001 ) and extended the latency period from 4 weeks in DMBA/TPA treatment group to 10 weeks in GOH-pretreated mice . Furthermore , we observed that GOH treatment significantly suppressed the Ras/Raf/ERK1/2 signaling pathway in skin tumor . Consistently , GOH-treated skin tumors showed reduced expression of Bcl-2 and increased expression of Bax in these lesions . Thus , it was concluded that GOH inhibits DMBA/TPA-mediated skin tumorigenesis by attenuating the Ras proliferation pathway and inducing pro-apoptotic state via inhibition of oxidative stress response and inflammation . OUTPUT: tumor promoting inflammation;resisting cell death INPUT: Comparative mutagenesis studies of N-(2'-deoxyguanosin-8-yl)-2-acetylaminofluorene ( dG-AAF ) and N-(2'-deoxyguanosin-8-yl)-2-aminofluorene ( dG-AF ) adducts positioned in the Nar I restriction enzyme site were performed using Escherichia coli ( E. coli ) and simian kidney ( COS-7 ) cells . Oligodeoxynucleotides ( (5)(')TCCTCG(1)G(2)CG(3)CCTCTC ) containing a recognition sequence for the Nar I restriction enzyme were modified site-specifically with dG-AAF or dG-AF . Modified and unmodified oligomers inserted into single-stranded phagemid shuttle vectors were used to transform E. coli or to transfect COS-7 cells . Following replication in host cells , progeny plasmids were recovered and analyzed for mutations . In SOS-induced E. coli , dG-AAF primarily induced one- and two-base deletions . The mutational frequency varied , depending on the position modified in the Nar I site ; 91% two-base deletions were observed at G(3) , while 8.4% and 2.8% deletions were detected at G(2) and G(1) , respectively . In contrast , dG-AF at any position in the Nar I site failed to produce deletions , generating primarily G --&gt ; T transversions ( mutational frequency , 7.6-8.4% ) . In COS-7 cells , both dG-AAF and dG-AF primarily induced G --&gt ; T transversions . Mutation frequencies for dG-AAF were 9.4-24% , the highest values being at G(1) and G(3) . Mutation frequencies for dG-AF were 9.3-21% , the higher value at G(2) . We conclude from this study that the mutation potential of dG-AAF and dG-AF depends on the structure of the adduct , the sequence context of the lesion , and the host cell used for the experiment . OUTPUT: genomic instability and mutation INPUT: Food-derived heterocyclic aromatic amines ( HCAs ) have proved to be carcinogenic in both rodents and nonhuman primates . Two different metabolic pathways are suggested for the metabolic activation of HCA . The hepatic pathway proceeds via a two-step process involving N-hydroxylation by cytochrome P4501A2 and subsequent O-acetylation by N-acetyltransferase-2 . An alternative pathway may be of particular interest in extrahepatic tissues and proceeds via one-electron oxidation catalyzed by prostaglandin H synthase ( PHS ) , rendering free-radical metabolites . In this study , we investigated the metabolic activation of two HCAs , 2-amino-3-methylimidazo[4,5-f]quinoline ( IQ ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , by two different enzyme systems in vitro , generating different primary and secondary reactive metabolites . Rat liver S9 mix and PHS were used as the activating system and represent the hepatic and extrahepatic pathways , respectively . Electron-spin resonance spectroscopy showed that both IQ and PhIP exerted inhibiting effects on PHS-mediated formation of hydroxyl radicals during the conversion of arachidonic acid to prostaglandins . Evidence for the formation of HCA free radicals was presented in an indirect way by the formation of glutathione-derived thiyl radicals , with purified PHS as the activating system . Activation by S9 mix did not result in the formation of detectable radical metabolites , showing that the two metabolic routes primarily led to the formation of different metabolites . In all electron-spin resonance experiments , IQ appeared to be more effective than PhIP . In contrasts , DNA adduct analysis by means of ( 32)P-postlabeling showed similar adduct patterns for S9 and PHS in single-stranded and double-stranded salmon testes DNA after incubation with PhIP , indicating the ultimate formation of a common reactive intermediate . For IQ , activation by PHS led to an additional adduct spot that was not present after S9 activation . Furthermore , activation of IQ resulted in higher adduct levels compared with PhIP for both activation pathways . Overall , adduct levels were higher in single-stranded DNA than double-stranded DNA . Our results showed that the hepatic and extrahepatic pathways resulted in different primary metabolites , while the ultimate formation of a similar reactive intermediate for PhIP , possibly an arylnitrenium ion , suggested that both pathways could play an important role in the onset of carcinogenesis . OUTPUT: genomic instability and mutation INPUT: Magnetite iron nanoparticles have been widely used as contrast agents and in thermal therapy for cancer . However , their adverse effects on human health have not been fully investigated . In this study , iron oxide nanoparticles were prepared using inorganic iron chloride ( size : 5.3+/-3.6 nm in phosphate buffered saline , surface charge : 23.14 mV ) , and their inflammatory responses were investigated . When mice were treated with iron oxide nanoparticles ( 250 microg/kg , 500 microg/kg , and 1mg/kg ) by a single intratracheal instillation , the level of intracellular reduced glutathione ( GSH ) was decreased in the cells of bronchoalveolar lavage ( BAL ) fluid . The arrest of cell cycles in G1 phase was observed , but S-phase was significantly decreased . The concentrations of pro-inflammatory cytokines ( IL-1 , TNF-alpha , and IL-6 ) were dose-dependently increased at day 1 after instillation in the BAL fluid and in the blood . During the experimental period of 28 days , pro-inflammatory cytokines ( IL-1 , TNF-alpha , and IL-6 ) , Th0 cytokine ( IL-2 ) , Th1 type cytokine ( IL-12 ) , Th2 type cytokines ( IL-4 and IL-5 ) , TGF-beta , and IgE were also elevated . Expressions of many genes related with inflammation or tissue damage such as heat shock protein , matrix metalloproteinase , tissue inhibitors of metalloproteinases , and serum amyloid A were significantly induced . Formation of microgranuloma , which is one of the indicators for chronic inflammatory response , was observed in the alveolar space . In addition , distribution of B cell and CD8+ T cell in blood lymphocytes was increased at day 28 . Based on the result , iron oxide nanoparticles may subchronic induce inflammatory responses via oxidative stress in mice by a single intratracheal instillation . OUTPUT: sustaining proliferative signaling;tumor promoting inflammation INPUT: This study was set up to investigate the relationships between the formation and removal of DNA damage in form of 8-oxodeoxyguanosine ( 8-oxodG ) in neonatal ( day 16 of gestation ) as compared to adult rats . The hypothesis addressed was whether the rapidly dividing foetal tissue has an enhanced requirement of DNA repair providing protection against potentially mutagenic DNA damages such as 8-oxodG . The activity of the primary 8-oxodG-repair protein OGG1 was measured by a DNA incision assay and the expression of OGG1 mRNA was measured by Real-Time PCR normalised to 18S rRNA . The tissue level of 8-oxodG was measured by HPLC-ECD . We found a 2-3-fold increased incision activity in the foetal control tissue , together with a 3-15-fold increase in mRNA of OGG1 as compared to liver tissue from adult rats . The levels of 8-oxodG in the foetal tissue were unaltered as compared to the adult groups . To increase the levels of 8-oxodG , the rats received an injection ( i.p. ) of the hepatotoxin 2-nitropropane . The compound induced significant levels of 8-oxodG in male rat livers 5h after the injection and in the foetuses 24h after the injection , while the female rats showed no increase in 8-oxodG . The incision activity was slightly depressed in both male and female liver tissue and in the foetal tissue 5h after the injection , but significantly increased from 5 to 24h after the injection . However , it did not reach levels significantly above the control levels . In conclusion , this study confirms that foetal tissue has increased levels of OGG1 mRNA and correspondingly an enhanced incision activity on an 8-oxodG substrate in a crude tissue extract . OUTPUT:

genomic instability and mutation;tumor promoting inflammation

HoC_dynamic_5_shot39

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Our previous study demonstrated that 5-aminolevulinic acid ( ALA ) administered to mice stimulates oxidative phosphorylation by upregulation of the mitochondrial respiratory chain complex IV enzyme cytochrome c oxidase ( COX ) . The present study investigated whether ALA disrupts the Warburg effect , which represents a shift in ATP generation from oxidative phosphorylation to glycolysis , protecting tumor cells against oxidative stress-mediated apoptosis . The human lung carcinoma cell line A549 exposed to ALA exhibited enhanced oxidative phosphorylation , which was indicated by an increase in COX protein expression and oxygen consumption . Furthermore , ALA suppressed glycolysis-mediated acidosis . This normalization of the ATP metabolic pathways significantly increased the generation of superoxide anion radical ( O2•- ) and the functional expression of active caspase-3 , leading to caspase-dependent apoptosis . These data demonstrate that ALA inhibits the Warburg effect and induces cancer cell death . Use of this endogenous compound might constitute a novel approach to cancer therapy . OUTPUT: cellular energetics;resisting cell death INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Focal inflammation causes systemic fever . Cancer hyperthermia therapy results in shrinkage of tumors by various mechanisms , including induction of adaptive immune response . However , the physiological meaning of systemic fever and mechanisms of tumor shrinkage by hyperthermia have not been completely understood . In this study , we investigated how heat shock influences the adaptive immune system . We established a cytotoxic T lymphocyte ( CTL ) clone ( #IM29 ) specific for survivin , one of the tumor-associated antigens ( TAAs ) , from survivin peptide-immunized cancer patients ' peripheral blood , and the CTL activities were investigated in several temperature conditions ( 37-41�C ) . Cytotoxicity and IFN-γ secretion of CTL were greatest under 39�C condition , whereas they were minimum under 41�C . To address the molecular mechanisms of this phenomenon , we investigated the apoptosis status of CTLs , expression of CD3 , CD8 , and TCRαβ by flow cytometry , and expression of perforin , granzyme B , and Fas ligand by western blot analysis . The expression of perforin and granzyme B were upregulated under temperature conditions of 39 and 41�C . On the other hand , CTL cell death was induced under 41�C condition with highest Caspase-3 activity . Therefore , the greatest cytotoxicity activity at 39�C might depend on upregulation of cytotoxic granule proteins including perforin and granzyme B. These results suggest that heat shock enhances effector phase of the adaptive immune system and promotes eradication of microbe and tumor cells . OUTPUT: resisting cell death INPUT: Photodynamic therapy ( PDT ) may trigger apoptosis or necrosis in cancer cells . Several steps in the induction and execution of apoptosis require high amounts of adenosine-5'-triphosphate ( ATP ) . Because the mitochondrial membrane potential ( delta psi ) decreases early in apoptosis , we raised the question about the mechanisms of maintaining a sufficiently high ATP level . We therefore monitored delta psi and the intracellular ATP level of apoptotic human epidermoid carcinoma cells ( A431 ) after photodynamic treatment with aluminum ( III ) phthalocyanine tetrasulfonate . A maximum of caspase-3-like activity and nuclear fragmentation was found at fluences of about 4 J cm(-2) . Under these conditions apoptotic cells reduced delta psi rapidly , while the ATP level remained high for 4-6 h after treatment for cells supplied with glucose . To analyze the contribution of glycolysis to the energy supply during apoptosis , experiments were carried out with cells deprived of glucose . These cells showed a rapid drop of ATP content and neither caspase activation nor nuclear fragmentation could be detected . We conclude that the use of glucose as a source of ATP is obligatory for the execution of PDT-induced apoptosis . OUTPUT:

resisting cell death;cellular energetics

HoC_dynamic_5_shot40

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Several studies have indicated that the cell-surface expressed nucleolin is implicated in tumorigenesis and angiogenesis , and represents an important target for cancer therapy . Here we show that treatment of rhabdoid tumor derived G401 cells with a nucleolin antagonist , the HB-19 pseudopeptide , could restore contact inhibition , impair anchorage-independent growth , and suppress tumor development in nude mice . G401 cells grow without contact inhibition , which is an in vitro characteristic property of malignant tumor cells . At concentrations of HB-19 that does not affect cell viability and multiplication index , there is restoration of contact inhibition thus suggesting that HB-19 treatment causes reversion of the malignant phenotype . Accordingly , HB-19 pretreated G401 cells lose the capacity to form colonies in soft agar . When assayed for tumorigenicity in nude mice , only 50% of mice injected with HB-19 pretreated G401 cells developed tumors with the mean tumor weight of 0.32 g , compared to 100% of mice injected with control G401 cells with the mean tumor weight of 2.36 g . Interestingly , the restoration of contact inhibition in HB-19 treated G401 cells is concomitant with marked reduction of transcripts coding the Wilms ' tumor 1 gene , matrix metalloproteinase-2 , epithelial isoform of CD44 , and vascular endothelial growth factor , whereas no apparent modification is detected for transcripts coding the proto-oncogene c-Myc , anti-apoptotic Bcl-2 , pro-apoptotic Bax , tissue inhibitor of metalloproteinase TIMP-1 , angiogenesis inhibitor TSP-1 , and growth factor Midkine . These findings indicate that the molecular mechanism of action of HB-19 on such highly malignant rhabdoid tumor cells is associated with a selective inhibitory effect on the expression of genes implicated in tumorigenesis and angiogenesis . OUTPUT: evading growth suppressors INPUT: Epidermal growth factor receptor-tyrosine kinase inhibitors ( EGFR-TKIs ) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor ( EGFR ) gene . On the other hand , some lung cancer patients with wild type EGFR also respond to EGFR-TKIs , suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells . However , the effect of EGFR-TKIs on host microenvironments is largely unknown . A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells . This model was used to investigate the therapeutic efficacy of erlotinib , an EGFR-TKI , on multiple organ metastases induced by human small cell lung cancer cells ( SBC-5 cells ) that did not express EGFR . Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro , it significantly suppressed bone and lung metastases in vivo , but not liver metastases . An immunohistochemical analysis revealed that , erlotinib significantly suppressed the number of osteoclasts in bone metastases , whereas no difference was seen in microvessel density . Moreover , erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line ( MC3T3-E1 cells ) . These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells . OUTPUT: activating invasion and metastasis;inducing angiogenesis;sustaining proliferative signaling INPUT: Cellular senescence is considered as a tumor suppressive mechanism . Recent evidence indicates however that senescent cells secrete various growth factors and cytokines , some of which may paradoxically promote cancer progression . This phenomenon termed senescence-associated secretory phenotype ( SASP ) must be inhibited in order for anti-proliferative agents to be effective . The present study was designed to determine whether the β-catenin destruction complex ( BCDC ) , known to integrate the action of various growth factors and cytokines , would represent a suitable target to inhibit the activity of SASP components . For this , we carried out experiments to determine the effect of drug-induced senescence on secretion of SASP , β-catenin transactivation , and the relationship between these processes . Moreover , genetic and pharmacological approaches were used to define the implication of BCDC in mediating the effects of SASP components on cell migration and resistance to drugs . The findings indicate that drug-induced senescence was associated with expression of various Wnt ligands in addition to previously known SASP components . Beta catenin transactivation and expression of genes implicated in epithelial-mesenchymal transition ( EMT ) also increased in response to drug-induced SASP . These effects were prevented by Pyrvinium , a recently described activator of BCDC . Pyrvinium also suppressed the effects of SASP on cell migration and resistance to doxorubicin . Together , these findings provide insights on the potential role of BCDC in mediating the effects of drug-induced SASP on cancer cell invasion and resistance to therapy , and suggest that targeting this pathway may represent an effective approach to enhance the activity of current and prospective anti-cancer therapeutics . OUTPUT: enabling replicative immortality;activating invasion and metastasis INPUT: p38 kinases activated by growth factors , hormones , and environmental stresses exert diverse functions in regulating normal and malignant cell pathophysiology . Enhanced levels of activated p38 isoforms have been linked with poor prognosis in breast cancer , although the mechanistic basis for this association is poorly understood . In this study , we report that p38 activation in cervical cancer cells is driven by osteopontin ( OPN ) , an extracellular matrix-associated cytokine that drives invasive progression . OPN regulates CD44-mediated p38 phosphorylation that induces NF-κB activation and NF-κB-dependent expression of furin , an extracellular protease implicated in human papilloma virus ( HPV ) processing that enhances cervical cancer cell motility . OPN induces CD44-mediated MKK3/6 phosphorylation which in turn phosphorylates p38 in these cells . OPN-induced furin expression and cell motility was impeded by blockades to MKK3/6 , p38α/β or NF-κB signaling . In a mouse xenograft model of human cervical cancer , tumor growth was enhanced by OPN overexpression and blocked by short hairpin RNA ( shRNA)-mediated OPN silencing . Furin overexpression similarly augmented tumor growth in the model , whereas blocking MKK3/6 , p38 , or furin reduced OPN-induced cervical tumor growth . Analysis of clinical specimens revealed that enhanced expression of OPN , phosphorylated NF-κB , p65 , and furin correlated with cervical cancer progression , further strengthening the in vitro and in vivo results . In summary , our findings offer a proof of concept for targeting OPN and its downstream p38 signaling as a novel therapeutic strategy to manage cervical cancer . OUTPUT: sustaining proliferative signaling INPUT: Biomaterials such as polyetherurethans ( PEUs ) are the scaffolding , which is indispensable for the development of the bio-artificial organs . However , PEUs can induce tumors in subcutaneous implantation sites in rat . We have shown that the different inhibitory potential of gap junctional intercellular communication ( GJIC ) on the surface of the biomaterials , including PEUs , is a key step in determining the tumorigenic potential . Here we show that suppression of a gap junctional protein connexin 43 ( Cx43 ) plays an important role in in vivo tumorigenesis induced by PEUs for the first time and that Cx43 transfection may be an effective strategy for preventing tumorigenesis induced by biomaterials . Rat tumor cell line U41 is derived from tumors in the subcutaneous implantation of PEU films . The GJIC and the expression of Cx43 were suppressed in U41 . The restoration of normal phenotype , such as reduction of growth rate , recovery of contact inhibition and loss of colony formation ability in soft agar , was achieved by Cx43 transfection . These results strongly suggest that suppression of Cx43 expression plays an important role in the development of rat malignant fibrous histiocytoma ( MFHC ) caused by PEUs and that Cx43 transfection is effective for prevention of tumorigenesis induced by PEUs . OUTPUT: evading growth suppressors INPUT: Recent studies have shown that the transcription factor , nuclear factor kappaB ( NF-kappaB ) , regulates critical survival pathways in a variety of different cell types , including human pancreatic cancer cells . The activation of NF-kappaB is controlled by proteasome-mediated degradation of its endogenous polypeptide inhibitor , inhibitor of nuclear factor kappaBalpha . We investigated the effects of PS-341 , a peptide boronate inhibitor of the proteasome in human pancreatic cancer cells in vitro and in vivo . Comparison of PS-341's effects on the growth of eight different human pancreatic cancer cell lines revealed marked heterogeneity in drug responsiveness , ranging from highly resistant ( IC50 &gt ; 10 microM ; Panc-48 , HS766T , and Mia-PaCa-2 ) to extremely sensitive ( IC50 &lt ; 40 nM ; L3.6pl , Hpaf2 , and BxPC3 ) . However , these effects did not correlate with differential inhibition of NF-kappaB activation . Direct quantification of apoptosis revealed that PS-341's effects on cell growth largely correlated with sensitivity to programmed cell death . Evaluation of PS-341's effects on established orthotopic tumor xenografts demonstrated that biweekly intravenous administration of the maximum-tolerated dose of the drug ( 1 mg/kg ) led to significant reductions in the volumes of L3.6pl tumors but not Mia-PaCa-2 tumors . Laser scanning cytometer-mediated quantification of drug-induced apoptosis in the xenografts confirmed that PS-341 induced DNA fragmentation and activation of caspase-3 in L3.6pl tumors but not in Mia-PaCa-2 tumors . However , histological examination of drug-treated tumors revealed extensive central necrosis and reductions in microvessel density and VEGF expression in both tumor types . Taken together , our results demonstrate that PS-341 inhibits the growth of human pancreatic tumors via direct effects on tumor cells and indirect effects on the tumor vasculature . OUTPUT:

resisting cell death;inducing angiogenesis

HoC_dynamic_5_shot41

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Chronic inflammation is a critical component in breast cancer progression . Pro-inflammatory mediators along with growth/survival factors within the tumor microenvironment potentiate the expression of pro-inflammatory cytokines ( IL-1 , IL-6 , TNF-α ) , chemotactic cytokines and their receptors ( CXCR4 , CXCL12 , CXCL8 ) and angiogenic factors ( VEGF ) that often overcome the effect of anti-inflammatory molecules ( IL-4 , IL-10 ) thus evading the host's antitumor immunity . Detailed knowledge , therefore , of the regulatory mechanisms determining cytokine levels is essential to understand the pathogenesis of breast cancer . HIF-1α and NF-κB transcription factors are important players for the establishment of a pro-inflammatory and potentially oncogenic environment . HIF-1α is the key mediator of the cellular response to oxygen deprivation and induces the expression of genes involved in survival and angiogenesis within solid hypoxic tumors . The expression of these genes is often modulated by the p53 tumor suppressor protein that induces apoptosis or cell cycle arrest in neoplastic cells . Functional crosstalk between HIF-1α and p53 pathways mediated by modulators shared between the two transcription factors such as SRC-1 and SIRT-1 differentially regulate the expression of distinct subsets of their target genes under variable stress conditions . In an attempt to shed light on the complex regulatory mechanisms involved in cancer-related inflammation , we investigated the role of the two common p53 and HIF-1α co-regulators SRC-1 and SIRT-1 , in the expression of the highly potent metastatic chemokine receptor CXCR4 . Both SRC-1 and SIRT-1 overexpression in DSFX-treated MCF-7 cells reduced CXCR4 cellular levels implying that both co-regulators are crucial factors in the determination of the metastatic potential of breast cancer cells . OUTPUT: activating invasion and metastasis INPUT: Targeting cancer cell metabolism is a new promising strategy to fight cancer . Metformin , a widely used antidiabetic agent , exerts antitumoral and antiproliferative action . In this study , the addition of metformin to 2-deoxyglucose ( 2DG ) inhibited mitochondrial respiration and glycolysis in prostate cancer cells leading to a severe depletion in ATP . The combination of the two drugs was much more harmful for cancer cells than the treatment with metformin or 2DG alone , leading to 96% inhibition of cell viability in LNCaP prostate cancer cells . In contrast , a moderate effect on cell viability was observed in normal prostate epithelial cells . At the cellular level , the combination of metformin and 2DG induced p53-dependent apoptosis via the energy sensor pathway AMP kinase , and the reexpression of a functional p53 in p53-deficient prostate cancer cells restored caspase-3 activity . In addition to apoptosis , the combination of metformin and 2DG arrested prostate cancer cells in G(2)-M . This G(2)-M arrest was independent of p53 and correlated with a stronger decrease in cell viability than obtained with either drug . Finally , metformin inhibited 2DG-induced autophagy , decreased beclin 1 expression , and triggered a switch from a survival process to cell death . Our study reinforces the growing interest of metabolic perturbators in cancer therapy and highlights the potential use of the combination of metformin and 2DG as an anticancerous treatment . OUTPUT: cellular energetics;resisting cell death INPUT: PURPOSE Pyruvate kinase isoenzyme M2 ( PKM2 ) is a key enzyme in aerobic glycolysis ; inhibition of PKM2 leads to the tumor growth inhibition . In this study , the effects of combined treatment with cisplatin ( DDP ) and a plasmid that expresses a short hairpin RNA ( shRNA ) targeting PKM2 on the growth of human A549 xenograft lung cancer model were investigated . METHODS The expression of PKM2 in A549 cells was determined by immunofluorescence . PKM2 expression levels were evaluated by Western blot analysis . In a human A549 lung cancer xenograft model , the effects of treatment with shRNA , with or without cisplatin , on tumor volume were determined . Apoptosis and cell proliferation status were examined to determine the mechanisms of tumor growth inhibition . RESULTS Expression of shRNA targeting PKM2 resulted in inhibition of PKM2 expression in A549 cells . In the lung cancer xenograft model , average tumor volume in the group treated with both cisplatin and shRNA was statistically lower than those of other groups ( P &lt ; 0.05 ) . The levels of apoptotic cells were significantly higher in samples from animals in the combined treatment group than those from untreated animals ( P &lt ; 0.05 ) . The cell proliferation rate , as determined by counting cells labeled with an anti-phospho-histone H3 , a marker for mitosis , was lower in samples from animals treated with both cisplatin and shRNA than in samples from other groups ( P &lt ; 0.05 ) . CONCLUSIONS Use of RNA interfering ( RNAi ) targeting PKM2 significantly inhibited tumor growth when combined with cisplatin in a human A549 lung cancer xenograft model . The enhanced antitumor activity of the combined treatment compared to treatment with shRNA alone may result in part from increased induction of apoptosis and augmented inhibition of cancer cell proliferation . OUTPUT: resisting cell death INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: p21 is a potent cyclin-dependent kinase inhibitor that plays a role in promoting G1 cell cycle arrest and cellular senescence . Consistent with this role , p21 is a downstream target of several tumour suppressors and oncogenes , and it is downregulated in the majority of tumours , including breast cancer . Here , we report that protein arginine methyltransferase 6 ( PRMT6 ) , a type I PRMT known to act as a transcriptional cofactor , directly represses the p21 promoter . PRMT6 knock-down ( KD ) results in a p21 derepression in breast cancer cells , which is p53-independent , and leads to cell cycle arrest , cellular senescence and reduced growth in soft agar assays and in severe combined immunodeficiency ( SCID ) mice for all the cancer lines examined . We finally show that bypassing the p21-mediated arrest rescues PRMT6 KD cells from senescence , and it restores their ability to grow on soft agar . We conclude that PRMT6 acts as an oncogene in breast cancer cells , promoting growth and preventing senescence , making it an attractive target for cancer therapy . OUTPUT: enabling replicative immortality;sustaining proliferative signaling INPUT: Like all cancers , breast cancer is considered to result in part from the accumulation of multiple genetic alterations leading to oncogene overexpression and tumor suppressor loss . More recently , CpG island hypermethylation is known to be associated with gene silencing in cancer , and these silenced genes can be reactivated by 5-aza-2'-deoxycytidine ( 5-Aza-CdR ) . Retionoic acid receptor beta 2 gene is a tumor suppressor gene and the chemopreventive effects of retinoids are due to induction of RAR beta 2 . In this study , the effect of 5-Aza-CdR RAR beta 2 restoration was investigated in the MRK-nu-1 human female breast cancer cell line . Changes of the RAR beta 2 methylation status were assessed by methylation-specific PCR . Reverse transcription PCR was used to evaluate RARb beta 2 restoration . Cell cycling and growth inhibition were studied using flow cytometric analysis of DNA content and CellTiter 96 AQueous non-radioactive cell proliferation assay , respectively. 5-Aza-CdR treatment resulted in complete demethylation of the RAR beta 2 gene . RAR beta 2 restoration was accompanied by cell cycle arrest ( increase in the G0/G1- and decrease in the S- and G2/M-phases ) and time-dependent growth inhibition . In conclusion , RAR beta 2 can be activated in vitro by 5-Aza-CdR , which may be one of the mechanisms for the tumor cell growth inhibition by 5-Aza-CdR . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot42

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: CONTEXT Papillary thyroid carcinoma ( PTC ) is the most frequent thyroid tumor and is responsible for the overall increase in thyroid cancer incidence . S100A11 ( calgizzarin ) , a member of the S100 Ca(2+)-binding protein family , is involved in several different biological processes . S100A11 has been found up-regulated in PTC , both at the mRNA and protein levels . OBJECTIVE Through a combination of expression analysis and functional in vitro and in vivo studies , we have attempted to gain insight into the relevance of S100A11 overexpression in PTC biology . DESIGN The expression of the S100A11 gene in PTC was investigated in several gene expression data sets . The effect of S100A11 silencing on the hallmarks of the malignant phenotype of several PTC-derived cell lines was investigated . In NIH3T3 cells , the cooperation of S100A11 with the different PTC-specific oncogenes was assessed . RESULTS We found that the S100A11 gene expression is frequently up-regulated in PTC , anaplastic thyroid carcinoma , but not in follicular thyroid carcinoma . S100A11 overexpression was also detected in PTC-derived cell lines , which were then used for functional studies . S100A11 silencing in PTC-derived cell lines did not affect cell proliferation , whereas it reduced the loss of contact inhibition , anchorage-independent growth , and resistance to anoikis . Cotransfection experiments in NIH3T3 cells showed that overexpression of the S100A11 gene was able to enhance the transforming capabilities of the different PTC-associated oncogenes by affecting the loss of contact inhibition , anchorage-independent growth , and in vivo tumor formation . CONCLUSION Our data indicate that S100A11 overexpression exerts a protumoral functional role in PTC pathogenesis . OUTPUT: evading growth suppressors INPUT: The polycomb group family protein BMI-1 is overexpressed by and functions as an oncogene in many different human cancers . We have previously shown that BMI-1 promotes the tumorigenicity of Ewing sarcoma family tumors ( ESFTs ) and that this is mediated independently of CDKN2A repression . In this study , we have discovered that high levels of BMI-1 confer resistance to contact inhibition in ESFT cells . Using stable retroviral transduction , we evaluated the consequences of BMI-1 knockdown on the growth of CDKN2A wild-type and mutant ESFT cells in subconfluent and confluent conditions . Although knockdown of BMI-1 had no effect on proliferation in low-density cultures , at high cell densities it resulted in cell cycle arrest and death . The normal cell contact inhibition response is mediated , in large part , by the recently described Hippo pathway which functions to inhibit cell proliferation and promote cell death by inactivating the Yes-Associated Protein ( YAP ) . Significantly , we found that YAP levels , activity and expression did not diminish in confluent ESFT cells that expressed high levels of BMI-1 . In contrast , YAP expression and nuclear localization were reduced in confluent BMI-1 knockdown cells suggesting that silencing of BMI-1 restored contact inhibition by restoring normal activation of the Hippo-YAP growth-suppressor pathway . Importantly , knockdown of YAP in ESFT cells resulted in profound inhibition of cell proliferation and anchorage-independent colony formation suggesting that stabilization and continued expression of YAP is critical for ESFT growth and tumorigenicity . Together , these studies reveal a previously unrecognized link between BMI-1 , contact inhibition and the Hippo-YAP pathway and suggest that resistance to contact inhibition in BMI-1 overexpressing cancer cells may be in part a result of Hippo inhibition and aberrant stabilization of YAP . OUTPUT: evading growth suppressors INPUT: Elevated insulin-like growth factor binding protein-related protein 1 ( IGFBP-rP1 ) mRNA in senescent human mammary epithelial cells suggested that the IGFBP-3 gene product may inhibit cell proliferation . To test this hypothesis , we used a retroviral vector to express IGFBP-rP1 cDNA in the IGFBP-rP1-deficient MCF-7 breast cancer cell line . Compared with control vector-transduced cells , cumulative cell numbers for IGFBP-rP1-transduced polyclonal or clonal cell cultures were reduced by 39 and 74% , respectively , after 1 week . Medium conditioned by IGFBP-rP1-producing cultures reduced cumulative cell numbers in parental MCF-7 cultures by 20% compared with medium from cultures of a control vector-transduced cell line . Nuclear fragmentation analysis and cell proliferation assays completed in the presence of the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone excluded apoptosis as the responsible mechanism . The percentage of cells containing senescence-associated beta-galactosidase activity was doubled compared with control cell cultures . Flow cytometry analysis indicated that twice as many noncycling cells were present in the IGFBP-rP1-transduced MCF-7 cell cultures compared with controls . These findings indicate that IGFBP-rP1 is an inhibitor of MCF-7 breast cancer cell proliferation and may act via a cellular senescence-like mechanism . OUTPUT: resisting cell death;enabling replicative immortality;sustaining proliferative signaling INPUT: We have already shown that IL-10 plays an important role in immunosuppression and metastatic dissemination in the rat B-cell lymphoma L-TACB model . It was suggested that the up-regulation of IL-10 production and IL-10 receptor ( IL-10R ) expression would be part of the transition from primary tumor to metastatic phenotype and that IL-10 , besides its immunosuppressive activity , may act as a growth factor for metastatic L-TACB cells . The treatment of L-TACB-bearing rats with a single low-dose cyclophosphamide decreased IL-10 production , reverted immunosuppression and induced the immunologic rejection of tumor metastasis without any effect on primary tumor growth . Our current aim was to investigate the effects of cyclophosphamide on the expression of IL-10 and IL-10R on primary and metastatic L-TACB cells . Considering that cyclophosphamide is a prodrug , we used mafosfamide , a compound that yields in vitro the same active metabolites as cyclophosphamide does in vivo . Mafosfamide induced down-regulation of IL-10 production and IL-10R expression on metastatic cells and , concomitantly , inhibited metastatic cell proliferation . We suggest that mafosfamide would inhibit the regulatory loop mediated by the IL-10/IL-10R system and , as a consequence , metastatic cell proliferation . These results may have a considerable impact on the design of new therapies for metastatic lymphomas . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: We have recently identified ICBP90 as being a protein able to bind in vitro a CCAAT box of the topoisomerase II alpha gene promoter . The aim of the present work was to check whether ICBP90 is able to regulate in vivo topoisomerase II alpha expression in human lung fibroblasts under various proliferating conditions . Transient transfection experiments performed on moderately growing human lung fibroblasts ( 50% of confluence ) showed that overexpression of ICBP90 is associated with an elevation of topoisomerase II alpha expression and an increase of the cell proliferation rate . In highly proliferating human lung fibroblasts ( 20% confluence ) overexpression of ICBP90 had no effect . In contrast , in non-proliferating fibroblasts ( 100% confluence ) overexpression of ICBP90 allowed recovery of topoisomerase II alpha expression levels with a concomitant overgrowth of confluent cell cultures . Our results show that ICBP90 regulates topoisomerase II alpha expression and is able to overcome cell contact inhibition signaling , suggesting that increased ICBP90 expression may be involved in carcinogenesis . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot43

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Mice defective in the mismatch repair ( MMR ) gene Msh2 manifest an enhanced predisposition to skin cancer associated with exposure to UVB radiation . This predisposition is further heightened if the mice are additionally defective for the nucleotide excision repair gene Xpc . To test the hypothesis that the predisposition of Msh2 mutant mice to skin cancer reflects a mutator phenotype associated with increased proliferation of skin cells following exposure to UV radiation , Msh2 mutant mice were exposed to the tumor promoter TPA . Such mice showed a robust proliferative response in the skin , but did not manifest evidence of dysplasia or neoplasia . We conclude that the predisposition of Msh2 mice to UVB radiation-induced skin cancer reflects an interaction between the processes of mismatch repair and some other excision repair mode , the exact nature of which remains to be established . OUTPUT: genomic instability and mutation INPUT: An aqueous extract of Kefir , fermented milk originally produced in the Caucasus mountains , suppressed morphological changes of human melanoma HMV-1 and SK-MEL cells and human normal fibroblastTIG-1 cells caused by UVC-irradiation , suggesting that UV damage can be suppressed by the Kefir extract . The addition of the Kefir extract after UVC-irradiation of HVM-1 cells resulted in a remarkable decrease in intracellular reactive oxygen species ( ROS ) which had been increased by UVC irradiation . The Kefir extract also stimulated unscheduled DNA synthesis and suppressed UVC-induced apoptosis of HMV-1 cells . A colony formation assay revealed that the Kefir extract rescued HMV-1 cells from cell death caused by UVC irradiation . The Kefir extract , as well as methyl methanethiosulfonate which is known to enhance the nucleotide excision repair ( NER ) activity , exhibited strong thymine dimer repair-enhancing activity . Epigalocatechin exhibited a weak NER activity but vitamins A , C , and E and catechin showed no NER activity . The thymine dimer repair-enhancing factors in the Kefir extract were heat-stable and assumed to be molecules with a molecular weight of less than 5000 . The treatment of HMV-1 cells with the Kefir extract during or before UVC- irradiation also prevented the generation of ROS and thymine dimmer , and suppressed the apoptosis of HMV-1 cells , suggesting that application of Kefir can prevent UV damage . OUTPUT: tumor promoting inflammation;genomic instability and mutation;resisting cell death INPUT: The DNA mismatch repair ( MMR ) pathway corrects specific types of DNA replication errors that affect microsatellites and thus is critical for maintaining genomic integrity . The genes of the MMR pathway are highly conserved across different organisms . Likewise , defective MMR function universally results in microsatellite instability ( MSI ) which is a hallmark of certain types of cancer associated with the Mendelian disorder hereditary nonpolyposis colorectal cancer . ( Lynch syndrome ) . To identify previously unrecognized deleted genes or loci that can lead to MSI , we developed a functional genomics screen utilizing a plasmid containing a microsatellite sequence that is a host spot for MSI mutations and the comprehensive homozygous diploid deletion mutant resource for Saccharomyces cerevisiae . This pool represents a collection of non-essential homozygous yeast diploid ( 2N ) mutants in which there are deletions for over four thousand yeast open reading frames ( ORFs ) . From our screen , we identified a deletion mutant strain of the PAU24 gene that leads to MSI . In a series of validation experiments , we determined that this PAU24 mutant strain had an increased MSI-specific mutation rate in comparison to the original background wildtype strain , other deletion mutants and comparable to a MMR mutant involving the MLH1 gene . Likewise , in yeast strains with a deletion of PAU24 , we identified specific de novo indel mutations that occurred within the targeted microsatellite used for this screen . OUTPUT: genomic instability and mutation INPUT: UVB from solar radiation is both an initiating and promoting agent for skin cancer . We have found that primary human keratinocytes undergo an apoptotic response to UVB . To determine whether these responses are altered during the course of immortalization , we examined markers of apoptosis in primary human foreskin keratinocytes ( HFK ) transduced with either a retroviral vector expressing the E6 and E7 genes of HPV-16 or with empty vector alone ( LXSN-HFK ) . Whereas LXSN-HFK as well as early passage keratinocytes expressing HPV-16 E6 and E7 ( p7 E6/7-HFK ) were both moderately responsive to UVB irradiation , late passage-immortalized keratinocytes ( p27 E6/7-HFK ) were exquisitely sensitive to UVB-induced apoptosis . After exposure to UVB , enhanced annexin V-positivity and internucleosomal DNA fragmentation were observed in p27 E6/7-HFK compared with either LXSN- or p7 E6/7-HFK . Caspase-3 fluorometric activity assays as well as immunoblot analysis with antibodies to caspase-3 and poly(ADP-ribose) polymerase revealed elevated caspase-3 activity and processing at lower UVB doses in p27 E6/7-HFK compared with LXSN- or p7 E6/7-HFK . In addition , the caspase inhibitor DEVD-CHO reduced the apoptotic response and increased survival of all three HFK types . Immunoblot analysis revealed that caspase-8 was activated in all three cell types , but caspase-9 was only activated in p27 E6/7-HFK . Cell cycle analysis further showed that only p27 E6/7-HFK exhibit G(2)/M accumulation that is enhanced by UVB treatment . This accumulation was associated with a rapid down-regulation of Bcl-2 in these cells . The immortalization process subsequent to the expression of HPV E6 and E7 may therefore determine UVB sensitivity by switching the mode of apoptosis from a caspase-8 to a Bcl-2-caspase-9-mediated pathway of apoptosis . OUTPUT: resisting cell death;enabling replicative immortality INPUT: Nitric oxide ( NO)-releasing non-steroidal anti-inflammatory drugs ( NO-NSAIDs ) which have been synthesized to reduce gastro-intestinal and cardiovascular toxicities of NSAIDs , possess anti-proliferative , pro-apoptotic and anti-cancer activities . Here , we show that NO-sulindac inhibited UVB-induced skin tumorigenesis in SKH-1 hairless mice . Topical application of NO-sulindac reduced tumor incidence , number ( p<0.05 ) and volume ( p<0.005 ) as compared to UVB ( alone)-irradiated vehicle-treated mice . An increase in TUNEL-positive cells in skin lesions was accompanied by the enhanced Bax:Bcl-2 ratio . The expression of pro-apoptotic Bax was increased whereas anti-apoptotic Bcl-2 reduced . However , proliferation was identified as the major target of NO-sulindac in this study . A reduced expression of PCNA and cyclin D1 associated with the dampening of cell cycle progression was observed . The mechanism of this inhibition was related to the reduction in UVB-induced Notch signaling pathway . UVB-induced inflammatory responses were diminished by NO-sulindac as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases Erk1/2 , p38 and JNK1/2 . In this regard , NO-sulindac also inhibited NFκB by enhancing IκBα as evidenced by the reduced expression of iNOS and COX-2 , the direct NFκB transcription target proteins . NO-sulindac significantly diminished the progression of benign lesions to invasive carcinomas by suppressing the tumor aggressiveness and retarding epithelial-mesenchymal transition . A marked decrease in the expression of mesenchymal markers such as Fibronectin , N-cadherin , SNAI , Slug and Twist and an increase in epithelial cell polarity marker E-cadherin were noted in NO-sulindac-treated tumors . Our data suggest that NO-sulindac is a potent inhibitor of UVB-induced skin carcinogenesis and acts by targeting proliferation-regulatory pathways . OUTPUT: resisting cell death;sustaining proliferative signaling;tumor promoting inflammation;activating invasion and metastasis INPUT: We have made xeroderma pigmentosum group A gene ( XPA)-knockout mice ( XPA(-/-) mice ) . The XPA(-/-) mice had no detectable activity for nucleotide excision repair ( NER ) and showed a high incidence of UVB-induced skin tumorigenesis . We have also found that cell lines derived from skin cancers in UVB-irradiated XPA(-/-) mice become tolerant to UV-irradiation and showed abnormal UV-induced cell cycle checkpoints and decreased mismatch repair ( MMR ) activity . These results suggested that the MMR-downregulation may help cells escape killing by UV-irradiation and thus MMR-deficient clones are selected for during the tumorigenic transformation of XPA(-/-) cells . In this report , we examined whether the incidence of UVB-induced skin tumorigenesis is enhanced in XPA(-/-)MSH2(-/-) , XPA(-/-) and MSH2(-/-) mice when compared with that in wild-type mice . Our results indicate that the MSH2-deficiency caused a high incidence of spontaneous and UVB-induced skin tumorigenesis and the XPA and MSH2 genes have additive roles in the UV-induced skin tumorigenesis . OUTPUT:

genomic instability and mutation;evading growth suppressors

HoC_dynamic_5_shot44

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: CDKN1C/P57 is a cyclin-dependent kinase inhibitor implicated in different human cancers , including hepatocellular carcinoma ( HCC ) ; however , little is known regarding the role of CDKN1C/P57 and its regulation in HCC . In this study , we show that the down-regulation of Notch1 and Notch3 in two HCC cell lines resulted in Hes1 down-regulation , CDKN1C/P57 up-regulation , and reduced cell growth . In line with these data , we report that CDKN1C/P57 is a target of transcriptional repression by the Notch effector , Hes1 . We found that the up-regulation of CDKN1C/P57 by cDNA transfection decreased tumor growth , as determined by growth curve , flow cytometry analysis , and cyclin D1 down-regulation , without affecting the apoptosis machinery . Indeed , the expression of Bax , Noxa , PUMA , BNIP(3) , and cleaved caspase-3 was not affected by CDKN1C/P57 induction . Morphologically CDKN1C/p57-induced HCC cells became flat and lengthened in shape , accumulated the senescence-associated β-galactosidase marker , and increased P16 protein expression . Evaluation of senescence in cells depleted both for Hes1 and CDKN1C/P57 revealed that the senescent state really depends on the accumulation of CDKN1C/p57 . Finally , we validated our in vitro results in primary HCCs , showing that Hes1 protein expression inversely correlates with CDKN1C/P57 mRNA levels . In addition , reduced Hes1 protein expression is accompanied by a shorter time to recurrence after curative resection , suggesting that Hes1 may represent a biomarker for prediction of patients with poor prognosis . OUTPUT: resisting cell death;enabling replicative immortality INPUT: Genotoxic stress induces cell cycle arrest and DNA repair which may enable tumor cells to survive radiation therapy . Here , we defined the role of Ca(2+) signaling in the cell cycle control and survival of chronic myeloid leukemia ( CML ) cells subjected to ionizing radiation ( IR ) . To this end , K562 erythroid leukemia cells were irradiated ( 0-10 Gy ) . Tumor survival was analyzed by clonogenic survival assay and cell cycle progression via flow cytometry . Plasma membrane cation conductance was assessed by patch-clamp whole-cell recording and the cytosolic free Ca(2+) concentration ( [ Ca(2+)](i) ) was measured by fura-2 Ca(2+) imaging . Nuclear activity of Ca(2+)/calmodulin-dependent kinase II ( CaMKII ) was defined by Western blotting . In addition , the effect of IR ( 5 Gy ) on the cation conductance of primary CML cells was determined . The results indicated that IR ( 10 Gy ) induced a G(2)/M cell cycle arrest of K562 cells within 24 h post-irradiation ( p.i. ) and decreased the clonogenic survival to 0.5 % of that of the control cells . In K562 cells , G(2)/M cell cycle arrest was preceded by activation of TRPV5/6-like nonselective cation channels in the plasma membrane 1-5 h p.i. , resulting in an elevated Ca(2+) entry as evident from fura-2 Ca(2+) imaging . Similarly , IR stimulated a Ca(2+)-permeable nonselective cation conductance in primary CML cells within 2-4 h p.i. . Ca(2+) entry , into K562 cells was paralleled by an IR-induced activation of nuclear CaMKII . The IR-stimulated accumulation in G(2) phase was delayed upon buffering [ Ca(2+)](i) with the Ca(2+) chelator BAPTA-AM or inhibiting CaMKII with KN93 ( 1 nM ) . In addition , KN93 decreased the clonogenic survival of irradiated cells but not of control cells . In conclusion , the data suggest that IR-stimulated cation channel activation , Ca(2+) entry and CaMKII activity participate in control of cell cycle progression and survival of irradiated CML cells . OUTPUT: evading growth suppressors INPUT: The immune system has an important role in tumor appearance and spreading . One of the most efficient subpopulations of cytotoxic cells in the destruction of tumors are NK cells . NK cells are activated and increase their cytotoxic potential and modulate their cytokine production after treatment with IFNgamma , IL-12 , TNFalpha and IL-2 . The investigation of the activity of NK cells was performed on peripheral blood lymphocytes ( PBL ) of 16 healthy controls and of 40 patients with metastatic breast carcinoma . Modulation of NK cells was performed with IL-2 , IL-7 , IL-12 , TNFalpha , monoclonal antibodies ( mAb ) for TNFalpha and TNFalpha receptors type I and II , as well as with sera of healthy controls and patients with breast cancer in different clinical stages . Modulating effect of the applied factors after in vitro treatment of PBL was evaluated by the cytotoxic assay using 51chromium . Our results indicate that IL-2 significantly increased the activity of NK cells of controls and breast cancer patients . The sera of patients with advanced breast cancer significantly reduced NK cell activity . IL-7 , IL-12 and mAb for TNFalpha do not significantly change the activity of NK cells . The presence of anti-TNFalpha mAb did not change the inhibitory effect of the sera of breast cancer patients with advanced disease on the activity of NK cells of controls and patients with breast cancer . Blocking of TNFalpha Rcs with mAbs decrease the reactivity of NK cells for IL-2 . The treatment of breast cancer patients with advanced clinical stage of breast cancer with IL-2 , as an additional therapy , could be advantageous , as NK cells after this treatment increase their cytotoxic activity against tumor cells and can improve therapeutical results . OUTPUT: avoiding immune destruction INPUT: Cellular senescence , initially observed during subculturing of normal diploid fibroblasts , can also be induced by chronic exposure to cellular stress , such as UV light , oxidative stress , or DNA damaging agents . Here we demonstrate that stable expression of an activated form of MKK6 ( MKK6EE ) , a direct activator of the stress-induced p38(HOG) mitogen-activated protein kinase pathway , is sufficient for inducing features of senescence including a flattened , vacuolated , and irregular morphology , staining for acidic beta-galactosidase , and accumulation of age-associated pigments . Consistent with the senescent phenotype , p38(HOG) activation induces a G(1) cell cycle arrest , which is permanent and irreversible after 4 days . MKK6EE also induces biochemical features of senescence in a p38-dependent manner , including enhanced expression of p21(CIP) , a cyclin-dependent kinase inhibitor . Microarray analysis of MKK6EE cells showed a pattern of gene expression noted previously in Werner Syndrome and senescent fibroblasts . These results define p38(HOG) as an intracellular pathway that activates a senescence checkpoint in tumor cells and may play a role in Ras- or stress-induced senescence . OUTPUT: enabling replicative immortality;sustaining proliferative signaling INPUT: Deregulation of D-type cyclin-dependent kinases ( CDK4 and 6 ) is widely observed in various human cancers , illustrating their importance in cell cycle control . Like other cyclin-dependent kinases ( CDKs ) , assembly with cyclins is the most critical step for activation of CDK4/6 . As previously reported elsewhere , we observed that the level of cyclinD1-CDK4 complex and its associated kinase activity were significantly low in asynchronously proliferating mouse embryo fibroblasts lacking both p21(Cip1) and p27(Kip1) ( p21/p27-null MEFs ) . These evidences imply that p21(Cip1) and p27(Kip1) CDK inhibitors are ' essential activators ' of cyclin D-kinases . We , however , discovered here that both the assembly and activation of cyclin D1-CDK4 complex occur when quiescent p21/p27-null MEFs were stimulated to re-enter the cell cycle . This mitogen-induced cyclin D1-kinase activity was blocked by overexpression of p16(INK4a) and resulted in the inhibition of S phase entry in p21/p27-null MEFs . Furthermore , ectopic expression of p34(SEI-1) , a mitogen-induced CDK4 binding protein , increased the levels of active cyclinD1-CDK4 complex in asynchronously proliferating p21/p27-null MEFs . Together , our results suggest that there are several independent ways to stimulate the assembly of cyclin D1-CDK4 kinases . Although p21(Cip1) and p27(Kip1) play a role in this process , our results demonstrate that additional mechanisms must occur in G0 to S phase transition . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Natural killer ( NK ) and CD56(+) T cells are thought to play a central role in antitumour immunity . Their cytolytic activities are controlled by a variety of receptors including CD94 and killer immunoglobulin-like receptors ( KIR ) , which bind to major histocompatibility complex ( MHC ) class I molecules on target cells and mediate cell activation or inhibition . We have examined the numbers , phenotypes and antitumour cytotoxic functions of hepatic NK and CD56(+) T cells isolated from 22 patients with hepatic malignancy and 19 healthy donors . Flow cytometry revealed that NK cell numbers were increased among hepatic mononuclear cells in malignancy compared to histologically normal livers ( mean : 38% vs 27% ; P=0.03 ) , but CD56(+) T cell numbers were not ( 28% vs 27% ) . NK cells and CD56(+) T cells from tumour-bearing livers exhibited lymphokine-activated killing of K562 targets and T cell receptor-mediated lysis of P815 cells . The expression of CD94 and the KIR isotypes CD158a , CD158b and KIR3DL1 by CD56(+) T cells and NK cells was significantly and consistently reduced in tumour-bearing livers compared to healthy livers ( P<0.05 in all cases ) . Simultaneous ligation of CD158a , CD158b and KIR3DL1 caused an overall partial inhibition of CD56(+) T cell cytotoxic activity , suggesting that the observed reductions in KIR(+) cell numbers in malignancy are likely to lead to enhanced cytotoxicity . Our results suggest that , while hepatic CD56(+) T cells are not expanded in malignancy , downregulation of KIR and CD94 expression may be a mechanism by which the hepatic immune system can be activated to facilitate tumour rejection . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot45

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: When the prostate cancer cells become unresponsive to androgen therapy , resistance to chemotherapy becomes imminent , resulting in high mortality . To combat this situation , lycopodine , a pharmacologically important bioactive component derived from Lycopodium clavatum spores , was tested against hormone sensitive ( LnCaP ) and refractory ( PC3 ) prostate cancer cells in vitro . This study aims to check if lycopodine has demonstrable anti-cancer effects and if it has , to find out the possible mechanism of its action . The MTT assay was performed to evaluate the cytotoxic effect . Depolarization of mitochondrial membrane potential , cell cycle , EGF receptor activity and apoptosis were recorded by FACS ; profiles of different anti- and pro-apoptotic genes and their products were studied by semi-quantitative RT-PCR , indirect-ELISA , western blotting . Drug-DNA interaction was determined by CD spectroscopy . Administration of lycopodine down-regulated the expression of 5-lipoxygenase and the 5-oxo-ETE receptor ( OXE receptor1 ) and EGF receptor , and caused up-regulation of cytochrome c with depolarization of mitochondrial inner membrane potential , without palpable change in p53 activity , resulting in apoptosis , cell arrest at G0/G1 stage and ultimately reduced proliferation of cancer cells ; concomitantly , there was externalization of phosphotidyl serine residues . CD spectroscopic analysis revealed intercalating property of lycopodine with DNA molecule , implicating its ability to block cellular DNA synthesis . The overall results suggest that lycopodine is a promising candidate suitable for therapeutic use as an anti-cancer drug . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: The relationship between thyroid hormone ( triiodothyronine , T(3) ) and breast cancer is unclear . We studied the effect of the c-erbA/TR alpha proto-oncogene encoding a functional T(3) receptor ( TR alpha 1 ) , of its ligand T(3) , and of its retroviral , mutated counterpart , the v-erbA oncogene , on the proliferation capacity of nontumorigenic mammary epithelial cells ( EpH4 ) . We found that EpH4 cells expressing ectopically TR ( EpH4 + TR alpha 1 ) or v-erbA ( EpH4 + v-erbA ) proliferated faster than parental EpH4 cells that contained low levels of endogenous TR . T(3) inhibited DNA synthesis and proliferation in EpH4 + TR alpha 1 cells but not EpH4 or EpH4 + v-erbA cells . The study of cell-cycle genes showed that T(3) decreased cyclin D1 RNA and protein levels in EpH4 + TR alpha 1 cells . In addition , T(3) downregulated the expression of T1 , a gene that is overexpressed in human breast adenocarcinomas and is induced by mitogens , serum , and several oncogenes and cytokines . Inhibition of the T1 gene by T(3) required both de novo mRNA and protein synthesis . Furthermore , T(3) abolished the induction of T1 by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate and inhibited the activity of an activation protein 1-dependent promoter ( -73-Col-CAT ) in EpH4 + TR alpha 1 cells , suggesting that interference with activation protein 1 transcription factor plays a part in the inhibition of the T1 gene . Our results showed that T(3) reduced the proliferation of mammary epithelial cells and inhibited the expression of cyclin D1 and T1 genes . OUTPUT: sustaining proliferative signaling INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: INTRODUCTION Natural herbal compounds with novel actions different from existing breast cancer ( BCa ) treatment modalities are attractive for improving therapeutic efficacy and safety . We have recently shown that penta-1,2,3,4,6-O-galloyl-β-D-glucose ( PGG ) induced S-phase arrest in prostate cancer ( PCa ) cells through inhibiting DNA replicative synthesis and G(1) arrest , in addition to inducing cell death at higher levels of exposure . We and others have shown that PGG through intraperitoneal ( i.p. ) injection exerts a strong in vivo growth suppression of human PCa xenograft models in athymic nude mice . This study aims to test the hypothesis that the novel targeting actions of PGG are applicable to BCa cells , especially those lacking proven druggable targets . METHODS Mono-layer cell culture models of p53-wild type estrogen receptor ( ER)-dependent MCF-7 BCa cells and p53-mutant ER-/progesterone receptor ( PR)- and Her2-regular ( triple-negative ) MDA-MB-231 BCa were exposed to PGG for a comprehensive investigation of cellular consequences and molecular targets/mediators . To test the in vivo efficacy , female athymic mice inoculated with MDA-MB-231 xenograft were treated with 20mg PGG/kg body weight by daily gavage starting 4 days after cancer cell inoculation . RESULTS Exposure to PGG induced S-phase arrest in both cell lines as indicated by the lack of 5-bromo2'-deoxy-uridine ( BrdU ) incorporation into S-phase cells as well as G(1) arrest . Higher levels of PGG induced more caspase-mediated apoptosis in MCF-7 , in strong association with induction of P53 Ser(15) phosphorylation , than in MDA-MB-231 cells . The cell cycle arrests were achieved without an induction of cyclin dependent kinase ( CDK ) inhibitory proteins P21(Cip1) and P27(Kip1) . PGG treatment led to decreased cyclin D1 in both cell lines and over-expressing cyclin D1 attenuated G(1) arrest and hastened S arrest . In serum-starvation synchronized MCF-7 cells , down-regulation of cyclin D1 was associated with de-phosphorylation of retinoblastoma ( Rb ) protein by PGG shortly before G(1)-S transition . In vivo , oral administration of PGG led to a greater than 60% inhibition of MDA-MB231 xenograft growth without adverse effect on host body weight . CONCLUSIONS Our in vitro and in vivo data support PGG as a potential drug candidate for breast cancer with novel targeting actions , especially for a triple negative BCa xenograft model . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: We examined the inducibility of drug resistance ( MDR1 , MRP1 , LRP ) and protein kinase C ( PKC ) isozyme ( alpha , epsilon , eta , theta , tau , zeta ) corresponding genes in A2780 ovarian cancer cells after a 24-hour treatment with adriamycin ( ADR ) , camptothecin ( CAM ) , etoposide ( ETO ) or vincristine ( VCR ) . Sublethal concentrations of drugs were used to exclude short-term effects caused by selection . Cell cycle analysis was performed to identify possible correlation between resistance factors , PKC isozymes and proliferation . We found a mostly combined induction of MDR1 , LRP , PKC tau and PKC zeta by CAM , ETO and VCR . PKC alpha , epsilon , eta and theta gene expression altered variably . Cell cycle analysis showed that A2780 cells responded with a marked G2/M arrest after a 24-hour treatment with CAM , ETO and VCR but an association between the induction of PKC isozymes corresponding genes and proliferation was not seen . Our analysis points to a possible link between atypical PKC tau/PKC zeta and MDR1/LRP in cytostatic stress response of cancer cells . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot46

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Prohibiting angiogenesis is an important therapeutic approach for fighting cancer and other angiogenic related diseases . Research focused on proteins that regulate abnormal angiogenesis has attracted intense interest in both academia and industry . Such proteins are able to target several angiogenic factors concurrently , thereby increasing the possibility of therapeutic success . Aquaporin-1 ( AQP1 ) is a water channel membrane protein that promotes tumour angiogenesis by allowing faster endothelial cell migration . In this study we test the hypothesis that AQP1 inhibition impairs tumour growth in a mouse model of melanoma . After validating the inhibitor efficacy of two different AQP1 specific siRNAs in cell cultures , RNA interference experiments were performed by intratumoural injections of AQP1 siRNAs in mice . After 6 days of treatment , AQP1 siRNA treated tumours showed a 75 % reduction in volume when compared to controls . AQP1 protein level , in AQP1 knockdown tumours , was around 75 % that of the controls and was associated with a significant 40 % reduced expression of the endothelial marker , Factor VIII . Immunofluorescence analysis of AQP1 siRNA treated tumours showed a significantly lower microvessel density . Time course experiments demonstrated that repeated injections of AQP1 siRNA over time are effective in sustaining the inhibition of tumour growth . Finally , we have confirmed the role of AQP1 in sustaining an active endothelium during angiogenesis and we have shown that AQP1 reduction causes an increase in VEGF levels . In conclusion , this study validates AQP1 as a pro-angiogenic protein , relevant for the therapy of cancer and other angiogenic-related diseases such as psoriasis , endometriosis , arthritis and atherosclerosis . OUTPUT: inducing angiogenesis INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: Colorectal cancer ( CRC ) arises as the consequence of progressive changes from normal epithelial cells through polyp to tumor , and thus is an useful model for studying metabolic shift . In the present study , we studied the metabolomic profiles using high analyte specific gas chromatography/mass spectrometry ( GC/MS ) and liquid chromatography tandem mass spectrometry ( LC/MS/MS ) to attain a systems-level view of the shift in metabolism in cells progressing along the path to CRC . Colonic tissues including tumor , polyps and adjacent matched normal mucosa from 26 patients with sporadic CRC from freshly isolated resections were used for this study . The metabolic profiles were obtained using GC/MS and LC/MS/MS . Our data suggest there was a distinct profile change of a wide range of metabolites from mucosa to tumor tissues . Various amino acids and lipids in the polyps and tumors were elevated , suggesting higher energy needs for increased cellular proliferation . In contrast , significant depletion of glucose and inositol in polyps revealed that glycolysis may be critical in early tumorigenesis . In addition , the accumulation of hypoxanthine and xanthine , and the decrease of uric acid concentration , suggest that the purine biosynthesis pathway could have been substituted by the salvage pathway in CRC . Further , there was a step-wise reduction of deoxycholic acid concentration from mucosa to tumors . It appears that to gain a growth advantage , cancer cells may adopt alternate metabolic pathways in tumorigenesis and this flexibility allows them to adapt and thrive in harsh environment . OUTPUT: cellular energetics INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Immunosuppression has been related to the incidence of tumor apparition , including endocrine tumors . The intrasplenic ovarian tumor ( luteoma ) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and , from the immunological perspective , is located in a critical organ involved in immune response . To establish if immunosuppression could alter the development of this experimental tumor , the effects of cyclosporin A ( CsA ) and dexamethasone ( Dex ) were evaluated . After surgery , tumor-bearing and sham animals were kept without treatment for 4 weeks ; thereafter , they were distributed into CsA ( 25 mg/kg ) , Dex ( 0.1 mg/kg ) , or vehicle ( 75:25 castor oil:ethanol ) groups and were injected on alternate days for 50 days . Body weight was evaluated weekly . Animals were sacrificed after a jugular vein blood sample was obtained . Thymi were weighed . Tumors were measured and placed in formaline for histological studies . Serum luteinizing hormone ( LH ) , follicle-stimulating hormone ( FSH ) , prolactin ( PRL ) , and estradiol were measured by radioimmunoassay . Hematological parameters were determined . CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals ( P &lt ; 0.01 ) . Dex significantly impaired weight increase in both groups of animals . CsA induced a significant weight loss in sham animals , not observed in tumor-bearing animals . Dex induced thymus weight loss in both groups , whereas CsA induced thymus weight loss only in sham animals . Only Dex induced a decrease in lymphocyte number in both groups . CsA induced an increase in monocyte number only in sham animals . Treatments did not alter LH , FSH , or estradiol , whereas PRL was increased by CsA only in sham rats . Neither Dex nor CsA induced any significant variations in tumor volume , nor did they alter tumor histology . In addition , no visible metastases or alterations in other organs were observed . We conclude that , though immunological parameters were altered by the treatments , immunosuppressor drugs did not condition tumor development . In addition , tumors secrete one or more factor/s that counteract CsA effect . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: Recent reports provide evidence that some growth factors behave as inhibitors of the apoptosis of the endothelial cells , bringing forward the concept of vascular survival as a post-angiogenesis process . At least two different vasculature development processes occur within a tumor : the angiogenic ( formation of new vessels ) and the vascular survival pathway , which is devoted to the preservation of the newly-formed vessels in layers that lose contact with the adjacent normal tissue . We developed a method to assess these processes in tissue samples . We noted that differences among tumors may exist not only in the tumor angiogenic activity ( TAA ) but also in the vascular survival ability ( VSA ) . One third of the highly angiogenic breast cancer cases examined had a poor ability to maintain high vessel density in inner tumor areas . Both parameters are independently related to prognosis , while VSA was directly related to tumor dimensions and node involvement . Patients with high TAA and VSA had a particularly poor prognosis . It is suggested that although cancer angiogenic activity is important for the local invasion and dissemination into vessels and lymphatics , the VSA may be important for the effective formation of viable tumor foci in lymph nodes or distant organs . Recognition and quantification of the vascular survival ability in human tumors may significantly improve the prognostic value of the assessment of tumor vasculature , and may help to stratify patients for clinical trials with novel anti-angiogenic or angiotoxic drugs . Elucidation of the pathways may provide additional targets for antiangiogenic therapy . OUTPUT:

inducing angiogenesis;activating invasion and metastasis

HoC_dynamic_5_shot47

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Immunosuppression has been related to the incidence of tumor apparition , including endocrine tumors . The intrasplenic ovarian tumor ( luteoma ) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and , from the immunological perspective , is located in a critical organ involved in immune response . To establish if immunosuppression could alter the development of this experimental tumor , the effects of cyclosporin A ( CsA ) and dexamethasone ( Dex ) were evaluated . After surgery , tumor-bearing and sham animals were kept without treatment for 4 weeks ; thereafter , they were distributed into CsA ( 25 mg/kg ) , Dex ( 0.1 mg/kg ) , or vehicle ( 75:25 castor oil:ethanol ) groups and were injected on alternate days for 50 days . Body weight was evaluated weekly . Animals were sacrificed after a jugular vein blood sample was obtained . Thymi were weighed . Tumors were measured and placed in formaline for histological studies . Serum luteinizing hormone ( LH ) , follicle-stimulating hormone ( FSH ) , prolactin ( PRL ) , and estradiol were measured by radioimmunoassay . Hematological parameters were determined . CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals ( P &lt ; 0.01 ) . Dex significantly impaired weight increase in both groups of animals . CsA induced a significant weight loss in sham animals , not observed in tumor-bearing animals . Dex induced thymus weight loss in both groups , whereas CsA induced thymus weight loss only in sham animals . Only Dex induced a decrease in lymphocyte number in both groups . CsA induced an increase in monocyte number only in sham animals . Treatments did not alter LH , FSH , or estradiol , whereas PRL was increased by CsA only in sham rats . Neither Dex nor CsA induced any significant variations in tumor volume , nor did they alter tumor histology . In addition , no visible metastases or alterations in other organs were observed . We conclude that , though immunological parameters were altered by the treatments , immunosuppressor drugs did not condition tumor development . In addition , tumors secrete one or more factor/s that counteract CsA effect . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers . A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation . We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products . This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine ( NMBzA ) , and the response that increased levels of dietary vitamin E has on this induced carcinogenesis . Female C57BL/6 mice received NMBzA or vehicle ( corn oil ) i.p. weekly for 3 weeks . Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet . As an assessment of free radical activity , exhaled ethane was measured prior to killing the animals at 26 weeks . Esophagi from the various mice groups were assessed for size and frequency of tumors . Livers homogenates were analyzed for vitamins A and E , lipid fluorescence , conjugated dienes and malondialdehyde . Hepatic levels of vitamin A and E were decreased ( P &lt ; 0.05 ) and indices of lipid peroxidation were greater ( P &lt ; 0.05 ) in NMBzA-treated mice relative to controls . Lipid peroxidation and serum transaminases ( ALT and AST ) were greatest in mice given NMBzA and infected with the retroviruses . Incidence of esophageal tumors were also greatest in the NMBzA-treated , immunocompromised animals . Mice fed vitamin E-supplemented diets showed increased ( P &lt ; 0.05 ) hepatic concentrations of vitamin E and vitamin A , decreased activities of serum transaminases , decreased indices of lipid peroxidation , and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice . These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals . OUTPUT: avoiding immune destruction INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: We report the formation , detection , quantitation and structural characterization of products resulting from the adduction of deoxynucleosides ( deoxyadenosine , deoxyguanosine , deoxycytidine and 5-methyldeoxycytidine ) to the catechol estrogens ( CE ) of estrone , estradiol-17beta and estradiol-17 alpha . The crude products are obtained in a one-pot synthesis through oxidation of catechols to quinones and subsequent Michael-type reaction with the deoxynucleosides in acidic medium . In all experiments , adducts are detected by electrospray ionization mass spectrometry analysis after HPLC separation ( LC/ESI/MS(n) ) . The two pyrimidines deoxycytidine and 5-methyldeoxycytidine yield only CE adducts to deoxynucleosides , which correspond to stable adducts on DNA . For purines , the results depend on the CE ( 2,3- or 3,4-catechols ) used , the function and configuration on carbon 17 ( ketone for estrone , alcohol for alpha and beta isomers of estradiol ) , and on the purine itself ( deoxyadenosine or deoxyguanosine ) . Both stable adducts and deglycosylated adducts are formed , and therefore formation of stable adducts on DNA as well as the loss of purines from the DNA strands could be possible . MS(2) and MS(3) experiments prove to be relevant for further structural determinations , enabling in some cases the elucidation of the regiochemistry of adduction on the A and B rings of the steroid moiety . OUTPUT: genomic instability and mutation INPUT: The effects of dose per fraction on the ability of amifostine exposure to elevate angiostatin levels in the serum of mice and to inhibit spontaneous metastases formation using the well-characterized murine Sa-NH sarcoma were investigated . Amifostine was administered intraperitoneally at doses of 50 , 100 , or 200 mg/kg every other day for 6 days to C3Hf/Kam mice until tumors reached an average size of 8 mm in diameter . Amifostine was again administered immediately following surgical removal of the tumor-bearing limbs by amputation , and then once more 2 days later . Nontumor-bearing control animals were treated using the same dosing and surgery schedules . The average number of pulmonary metastases per animal was determined for each experimental group . A significant reduction ( P <.05 ) in the average number of pulmonary metastases was observed only in the group of animals exposed to a dose per fraction of 50 mg/kg . A dose of 100 mg/kg was less effective while 200 mg/kg had no effect on metastases formation in this study . The effects of amifostine exposure on serum levels of the angiogenesis inhibitor angiostatin were also determined using Western analysis . Correlating with the antimetastatic effect measured , exposure of animals to 50 mg/kg of amifostine resulted in a four-fold enhanced serum level of angiostatin above control levels . This phenomenon occurred in both tumor-bearing as well as nontumor-bearing animals . In contrast , a dose of 200-mg/kg amifostine administered intraperitoneally under these conditions had no measurable effect on angiostatin serum levels in this animal system . The enhanced ability of relatively low doses of amifostine to inhibit spontaneous metastases formation suggests that effective antimetastatic therapies with amifostine can be designed with minimal toxic side effects . While the dose responses for angiostatin production and metastases inhibition by amifostine are well correlated , the precise mechanism of action underlying these phenomena is unclear but is suggestive of a redox driven process(es) . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot48

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: INTRODUCTION Tamoxifen is the most widely prescribed anti-estrogen treatment for patients with estrogen receptor ( ER)-positive breast cancer . However , there is still a need for biomarkers that reliably predict endocrine sensitivity in breast cancers and these may well be expressed in a dynamic manner . METHODS In this study we assessed gene expression changes at multiple time points ( days 1 , 2 , 4 , 7 , 14 ) after tamoxifen treatment in the ER-positive ZR-75-1 xenograft model that displays significant changes in apoptosis , proliferation and angiogenesis within 2 days of therapy . RESULTS Hierarchical clustering identified six time-related gene expression patterns , which separated into three groups : two with early/transient responses , two with continuous/late responses and two with variable response patterns . The early/transient response represented reductions in many genes that are involved in cell cycle and proliferation ( e.g . BUB1B , CCNA2 , CDKN3 , MKI67 , UBE2C ) , whereas the continuous/late changed genes represented the more classical estrogen response genes ( e.g . TFF1 , TFF3 , IGFBP5 ) . Genes and the proteins they encode were confirmed to have similar temporal patterns of expression in vitro and in vivo and correlated with reduction in tumour volume in primary breast cancer . The profiles of genes that were most differentially expressed on days 2 , 4 and 7 following treatment were able to predict prognosis , whereas those most changed on days 1 and 14 were not , in four tamoxifen treated datasets representing a total of 404 patients . CONCLUSIONS Both early/transient/proliferation response genes and continuous/late/estrogen-response genes are able to predict prognosis of primary breast tumours in a dynamic manner . Temporal expression of therapy-response genes is clearly an important factor in characterising the response to endocrine therapy in breast tumours which has significant implications for the timing of biopsies in neoadjuvant biomarker studies . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Immunosuppression has been related to the incidence of tumor apparition , including endocrine tumors . The intrasplenic ovarian tumor ( luteoma ) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and , from the immunological perspective , is located in a critical organ involved in immune response . To establish if immunosuppression could alter the development of this experimental tumor , the effects of cyclosporin A ( CsA ) and dexamethasone ( Dex ) were evaluated . After surgery , tumor-bearing and sham animals were kept without treatment for 4 weeks ; thereafter , they were distributed into CsA ( 25 mg/kg ) , Dex ( 0.1 mg/kg ) , or vehicle ( 75:25 castor oil:ethanol ) groups and were injected on alternate days for 50 days . Body weight was evaluated weekly . Animals were sacrificed after a jugular vein blood sample was obtained . Thymi were weighed . Tumors were measured and placed in formaline for histological studies . Serum luteinizing hormone ( LH ) , follicle-stimulating hormone ( FSH ) , prolactin ( PRL ) , and estradiol were measured by radioimmunoassay . Hematological parameters were determined . CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals ( P &lt ; 0.01 ) . Dex significantly impaired weight increase in both groups of animals . CsA induced a significant weight loss in sham animals , not observed in tumor-bearing animals . Dex induced thymus weight loss in both groups , whereas CsA induced thymus weight loss only in sham animals . Only Dex induced a decrease in lymphocyte number in both groups . CsA induced an increase in monocyte number only in sham animals . Treatments did not alter LH , FSH , or estradiol , whereas PRL was increased by CsA only in sham rats . Neither Dex nor CsA induced any significant variations in tumor volume , nor did they alter tumor histology . In addition , no visible metastases or alterations in other organs were observed . We conclude that , though immunological parameters were altered by the treatments , immunosuppressor drugs did not condition tumor development . In addition , tumors secrete one or more factor/s that counteract CsA effect . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: Spindle cell oncocytoma ( SCO ) of the pituitary gland is a relatively recently established , very rare subtype of adenohypophysis tumours that was introduced as a distinct clinicopathological entity in the fourth edition of WHO classification of the central nervous system tumours ( 2007 ) . It is non-endocrine neoplasm of the anterior pituitary that occurs in adults and usually follows a benign clinical course , corresponding to WHO grade I. Up to now , pituitary SCO have been reported occasionally and only 14 cases of SCO have been documented in the literature . Because of their rarity , the pathogenesis and natural history of these tumours have not been fully characterized . We report two additional cases of SCO occurring in females aged 63 years ( Case 1 ) and 65 years ( Case 2 ) , who presented with pan-hypopituitarism , headache and visual field defect . In both cases , the magnetic resonance imaging showed solid sellar mass of moderate size with suprasellar extension . The clinical and radiological features suggested non-functioning pituitary macroadenomas without evidence of invasive growth . One patient presented with tumour recurrence 3 years after undergoing the previous surgical removal of tumour , which was initially misdiagnosed as schwannoma . The first tumour was removed by transsphenoidal surgery and the second one by frontal craniotomy . Histologically and immunohistochemically , both tumours displayed the features typical for SCO of the pituitary . They were composed of interwoven fascicles of spindle cells exhibiting abundant eosinophilic cytoplasm of oncocytic or granular appearance . Mitoses were rarely observed and necrosis was absent . In one case , the advanced lymphocytic infliltration was observed within neoplastic tissue . The tumour cells exhibited immunoreactivity for S-100 protein , galectin-3 , vimentin and epithelial membrane antigen but they were negative for GFAP , anterior pituitary neuroendocrine markers ( prolactin , growth hormone , TSH , ACTH , FSH , LH ) , chromogranin , synaptophysin , cytokeratin CK ( AE1/AE3 ) , smooth muscle actin , desmin , CD34 and CD68 . MIB1 labeling index did not exceed 10% . Ultrastructurally , the tumour cells were rich in mitochondria with lamellar cristae . Moreover , in Case 2 some tumour cells showed a number of giant mitochondria with severely destructed internal matrix . Spindle cell oncocytoma of the anterior pituitary is often misdiagnosed entity of uncertain histogenesis . It should be considered in the differential diagnosis of various sellar-region lesions of oncocytic morphology . OUTPUT: resisting cell death INPUT: Among salivary gland neoplasms are a group of rare tumors that are histologically identical to benign mixed tumors that inexplicably metastasize ; they have been called metastasizing mixed tumor ( MZMT ) of salivary glands . We report the clinicopathologic features and flow cytometric findings for 11 cases of MZMT . At the time of discovery of metastatic disease , the patients , six women and five men , ranged in age from 20 to 83 years . Primary sites of involvement included the parotid gland ( eight cases ) , submandibular gland ( two cases ) , and the nasal septum ( one case ) . With one exception , all the patients had at least a single recurrences of their primary mixed tumor , but two or more recurrences were the norm before development of metastatic foci . The metastases were discovered from six to 52 years following the occurrence of the primary tumor . Metastatic deposits were identified in bone , lung , regional lymph nodes , skin , kidney , retroperitoneum , oral cavity , pharynx , calvarium , and central nervous system . The metastases either occurred simultaneously with an episode of recurrent mixed tumor ( n = 5 ) or from 5 to 29 years after a recurrence ( n = 6 ) . The treatment of the primary , recurrent , and metastatic neoplasms was surgical excision . Follow-up , ranging from 8 months to 16 years following the diagnosis of MZMT , revealed seven patients to be alive without disease ( 64% ) and two dead of causes unrelated to metastatic disease ( 18% ) . Two patients ( 18% ) died as a direct result of metastatic tumor at 3 and 2 years after metastasis of their mixed tumors . Flow cytometric analysis revealed a diploid DNA cell population in the primary and/or metastatic tumors in nine cases . Aneuploid DNA cell content was identified in two of the cases . DNA ploidy levels and cell proliferation rates were compared with those of conventional benign mixed tumors and also with malignant mixed tumors . Retrospective analysis of histologic parameters ( mitotic rate , cellular pleomorphism , infiltrative growth , vascular or lymphatic invasion ) and flow cytometric analysis failed to identify criteria to predict the development of metastasis in these neoplasms . OUTPUT: activating invasion and metastasis;genomic instability and mutation INPUT: AIM To develop a simple , fast and inexpensive approach as well as an instrument for detection of gene mutation . METHODS Pyrosequencing based on bioluminometry assay was employed to detect gene mutation . Pyrosequencing is a method of sequencing by synthesis step-by-step using four enzymes , DNA-polymerase , ATP sulfurylase , luciferase and apyrase . The signal was produced by detecting pyrophosphate released during a dNTP incorporation . For mutation detection , a DNA fragment was amplified by PCR at first , followed by a single-stranded DNA preparation . In the second step , a short primer was annealed to the position just before the mutation point . Finally , specific dNTPs were added in terms of the template sequence . The mutation species can be readily determined by the sequence . RESULTS A new instrument was developed for gene mutation detection by pyrosequencing . To iteratively inject small amount of each dNTP for the sequencing reaction , capillaries were used to connect dNTP reservoirs and the reaction chamber . Each dNTP was delivered by adding a gas pressure on the top of a corresponding dNTP reservoir , by which 0.2 microL of dNTP can be exactly added each time . It was theoretically proved that undesired liquid seep through the capillary did not affect the sequencing reactions in pyrosequencing . In addition , the three possible variants ( wildtype , mutant and heterozygote ) of a mutant point Cys275Ser in P53 gene exon 8 were determined by pyrosequencing using the instrument . A simple method was also described for rapidly distinguishing the type of a variant . CONCLUSION The developed method is very simple , and the corresponding instrument is inexpensive and easy to operate , which can be used to detect many types of mutation . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot49

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: OBJECTIVE To construct the eukaryotic expression vector for Max interacting protein 1 ( Mxi1 ) . METHODS The full length cDNA of Mxi1 gene obtained from fetal lymphocyte and KG1 cells were inserted into plasmid pDs-red2-N1 respectively to generate pDs-red2-N1/Mri1 ( wild/mutation type ) . Then the recombinant vector was transfected into Cos-7 cells via liposome. 48 hours post transfection , mRNA of Mri1 gene was detected by RT-PCR and Mxi1 protein expression was detected by flow cytometry and fluorescence microscope in the Cos-7 cells . RESULTS The eukaryotic expression vector of Mxi1 was constructed and transfected into eukaryotic cells successfully . The expression of red fluorescence protein in the transfected Cos-7 cells was observed under fluorescence microscope which implied the expression of Mxi1 . The transfect efficiency of both wild and mutation type were in a high level in 3 days after transfected , which lasted to 6 d . RT-PCR amplified the total RNA extracted from the transfected Cos-7 cells could find increased mRNA level of Mxi1 gene . CONCLUSION We successfully constructed the eukaryote expression vector for Mri1 gene ; Cos-7 cells transfected with the vector via liposome could express Mxi1 protein . These could be useful for the further study of the Myc gene modulation . OUTPUT: genomic instability and mutation INPUT: BACKGROUND Adenosine receptors ( A1 , A2A , A2B , A3 ) play an important role in the regulation of growth , proliferation and death of cancer and normal cells . We recently showed the expression profile of A2A and A2B receptors in normal and tumor breast tissues . In the present study , we used semiquantitative RT-PCR to measure the A1 and A3 gene expression levels in normal and tumor breast tissues . METHODS Breast tumors ( n = 18 ) and non-neoplastic mammary tissues ( n = 10 ) were collected and histologically confirmed to be neoplastic or non-neoplastic , respectively . Total RNA was extracted and reverse transcribed into cDNA , and PCR was performed under optimized condition for each receptor subtype . Amplification of beta-actin mRNA served as control for RT-PCR . The PCR products were separated on 1.7% agarose gels . The intensity of the bands was quantitated with ImageJ software after normalization against beta-actin expression . RESULTS All breast tumor and normal tissue specimens expressed A1 and A3 adenosine receptor transcripts . However , we observed that the expression level of the A3 receptor in tumor tissues was 1.27-fold that of normal tissues , whereas there was no significant difference between the expression levels of A1 in normal and tumor tissues . CONCLUSIONS Interestingly , the results of the present study indicate that breast tumors exhibit a higher level of A3 transcripts ( than normal tissues ) and support the possible key role of A3 adenosine receptor in tumor development . However , further studies based on real-time quantitative RT-PCR are needed to identify the exact gene expression levels . OUTPUT: sustaining proliferative signaling INPUT: OBJECTIVE To observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe ( RSR ) on HER2 positive breast cancer , to evaluate the effects of RSR on the expressions of matrix metalloproteinases ( MMPs ) and the tissue inhibitor of metalloproteinases ( TIMPs ) in the recurrence and metastasis of HER2 positive breast cancer , thus revealing its anti-tumor recurrent and metastatic mechanisms . METHODS Selected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu . The primary tumor resection was carried out . After surgery they were randomly divided into the blank control group , the RSR group , the Herceptin group , and the combination group ( RSR + Herceptin group ) . The treatment lasted for 4 months . The inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated . The expressions of matrix metalloproteinase-2 ( MMP-2 ) , matrix metalloproteinase-9 ( MMP-9 ) , tissue inhibitor of metalloproteinase ( TIMP-1 ) , and TIMP-2 in the recurrent tumor tissue were detected using Western blot . RESULTS By the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group , showing statistical difference between the two groups ( P = 0.037 ) . The average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group . The inhibition rate of lung metastasis was 37. 85% in the RSR group , but with no statistical significance . The expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group , the Herceptin group , and the combination group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group , the Herceptin group , and the combination group was significantly down-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group ( P &lt ; 0.05 ) . The expressions of both TIMP-1 and TIMP-2 of the RSR group , the Herceptin group , and the combination group were all up-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group ( P &lt ; 0.05 ) . It was higher in the combination group than in the RSR group ( P &lt ; 0.05 ) . CONCLUSIONS RSR could inhibit the tumor recurrence of FVB/neu mice . It could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix . It might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells . OUTPUT: activating invasion and metastasis INPUT: This study intended to investigate the expression of the ZEB1 and E-cadherin proteins in lung squamous cell carcinoma ( LSCC ) tissues and to examine the clinicopathological correlation between protein levels and LSCC . RT-PCR and Western blot were used to examine the expression of ZEB1 and E-cadherin mRNAs and proteins in LSCC tissues as well as in adjacent normal tissues , and then analyze the relationship between the clinicopathological characteristics and the expression changes of ZEB1 and E-cadherin mRNAs in LSCC . In addition , RNAi was used to knockdown the expression of the ZEB1 gene in Human HCC827 cells ; subsequently , changes in the invasive ability of the resultant cells were studied . The positive rates of ZEB1 and E-cadherin mRNAs in LSCC tissues were 69.2 and 38.5% , respectively . They differed significantly from the corresponding positive rates in the adjacent normal lung tissues ( 15.4 and 80.8% , p<0.05 ) . There was a negative correlation between the protein levels of ZEB1 and E-cadherin in LSCC tissues ( r=-0.714 , p<0.001 ) ; in addition , it was found that ZEB1 protein expression in LSCC tissues was significantly higher than that in the neighboring normal lung tissues ( p<0.05 ) , and its expression was also significantly higher in patients with lymph node metastases and distant metastases compared to those patients without metastatic disease ( p<0.05 ) . On the contrary , E-cadherin expression was significantly lower in LSCC tissues than that in the neighboring normal tissue ( p<0.05 ) . It was lower in patients with lymph node metastasis and distant metastasis compared to patients without metastatic disease ( p<0.05 ) . However , the expression of ZEB1 and E-cadherin was independent of gender , age , tumor size , or tumor differentiation level ( p>0.05 ) . Transfection of ZEB1 siRNA into HCC827 cells significantly reduced the ZEB1 protein level ( p<0.01 ) and significantly elevated E-cadherin levels ( p<0.01 ) . Moreover , significantly less ZEB1 siRNA-transfected cells migrated through Transwell chambers in the LSCC tissue than that in the control groups ( untransfected or transfected with control siRNA , p<0.01 ) . The expression of the ZEB1 gene in LSCC tissues is downregulated with the expression of E-cadherin . On the other hand , the expression of siRNA against ZEB1 promotes E-cadherin expression and suppresses the invasive ability conferred by E-cadherin . In conclusion , our data suggested that overexpression of the ZEB1 gene is possibly associated with the occurrence , development , invasion of LSCC . OUTPUT: activating invasion and metastasis INPUT: c-myc , c-erbB-2 , and Ki-67 expression was examined by immunohistochemistry in 11 normal breast tissues and 42 invasive and 14 noninvasive breast carcinomas . The c-myc product was detected in all breast carcinoma specimens and in 7 of 11 normal breast tissues . Invasive tumors stained more frequently with the anti-myc monoclonal antibody than did noninvasive tumors , while the level of expression in normal breast tissue was much less than that in breast cancer . Membrane staining of the c-erbB-2 protein was demonstrated in 29% ( 4 of 14 ) of noninvasive ductal carcinomas and in 45% ( 19 of 42 ) of invasive breast carcinomas . None of the 11 normal breast tissue samples was positive . The mean value of Ki-67-positive cells was 0.91 +/- 0.31% for normal breast tissue , 4.57 +/- 1.36% for noninvasive ductal carcinoma , and 12.76 +/- 2.18% for invasive breast cancer . In 42 invasive breast carcinomas , the expression of c-myc , c-erbB-2 , and Ki-67 proliferation marker were compared with lymph node status , estrogen receptor status , progesterone receptor status , and age of patients at diagnosis. c-erbB-2 overexpression and Ki-67 overexpression were identified as the only factors associated with lymph node status . We concluded that they might be additional prognostic factors for breast carcinoma . OUTPUT: sustaining proliferative signaling INPUT: OBJECTIVE To investigate the expression and mutation of MTA1 , nm23H1 and E-cadherin(E-cad) genes in ovarian carcinoma ( OC ) in relation to lymph node ( LN ) metastasis . METHODS A panel of normal ovarian tissues , primary OC specimens and corresponding LNS was examined for mRNA expression and mutation of MTA1 and nm23H1 and protin expression of E-cad genes by using RT-PCR , RT-PCR-SSCP and immunohistochemistry . RESULTS The frequency of MTA1 over expression was 100%(7/7) in primary OC with metastasis but only 38.5%(5/13) in those without metastasis ( P = 0.0103 ) . Overexpression of MTA1 was observed in 87.5%(6/7) of LNS with metastasis but in only 23%(3/13) of LNS without metastasis ( P = 0.0118 ) . In contrast with MTA1 , low expression of nm23H1 mRNA was seen in 7 of 7 OC with metastasis but only in 4 of 13(30%) of those without metastasis ( P = 0.0043 ) . Low nm23H1 expression was also seen in 7 of 7 LNS with metastasis but only in 5 of 13 ( 38.5% ) nonmetastatic LNS ( P = 0.0102 ) . Meantime , no expression of E-cad protein was observed in 7 of 7 OC with metastasis but in 6 of 13(46.2%) of those without metastasis ( P = 0.044 ) . In correlation analysis of the three genes , MTA1 reversely correlated with nm23H1 and E-cad respectively ( r = -0.903 , -0.803 ) , and positive correlation existed between nm23H1 and E-cad ( r = 0.724 ) . No mutation of MTA1 , nm23H1 and was found by SSCP analysis . CONCLUSION The mRNA expression of MTA1 , nm23H1 and E-cad is positively and negatively correlated with LN metastasis . The expression abnormalities but not the mutations of the three genes are frequent events related to LN metastasis of ovarian cancer . OUTPUT:

activating invasion and metastasis;genomic instability and mutation

HoC_dynamic_5_shot50

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND : CD81 is a transmembrane protein that serves as a putative receptor for hepatitis C virus . In addition , CD81 has been suggested to be involved in a broad range of other cellular functions . Its putative implication in tumorigenesis has so far , however , remained largely unexplored . To assess the candidacy of CD81 as a tumor suppressor in gastric cancer development , we investigated its expression and function in a series of primary gastric tumors and gastric tumor-derived cell lines . METHODS : The expression and concomitant methylation status of the CD81 gene and its effect on tumor development and cellular signaling were evaluated . RESULTS : CD81 mRNA levels were found to be low in 16 of 40 ( 40% ) primary tumors and 9 of 14 ( 64.2% ) cell lines , and these low expression levels were found to correlate with the stage and grade of the tumors . Genomic alterations of CD81 were not encountered , whereas its expression could be re-activated in low expressing cells upon 5-aza-dC treatment . Bisulfite DNA sequencing analysis of 10 CpG sites within the 5 ' proximal region of the CD81 gene promoter revealed that the observed transcriptional silencing was tightly associated with aberrant hypermethylation . Subsequent restoration of CD81 expression induced a G(1) cell cycle arrest and apoptosis , whereas siRNA-mediated CD81 down-regulation promoted cell proliferation and attenuated cellular responses to various apoptotic stress stimuli . Also the colony-forming ability of the tumor cells could be inhibited and enhanced through CD81 up- and down-regulation , respectively . CD81 was found to inhibit p38 ( but not ERK , JNK and AKT ) phosphorylation and its growth suppressive effect could be abolished through p38 up- and down-regulation . CONCLUSION : From our data we conclude that epigenetic inactivation of CD81 is a common feature of gastric tumors and that this inactivation may render growth and survival advantages to the tumor cells , at least partially through p38 signaling . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: Zinc dyshomeostasis can induce cell death . However , the mechanisms involved have not been fully elucidated in prostate cancer ( PCa ) cells , which differ dramatically from normal cells in their zinc handling ability . Here , we studied the effects of the ionophore Zn-pyrithione ( ZP ) and the chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine ( TPEN ) . Both compounds induced cell death at micromolar concentrations when incubated with androgen-dependent ( LNCaP ) , androgen-independent ( PC3 , DU145 ) and androgen-sensitive ( C4-2 ) PCa cell-lines . Compared to PCa cells , RWPE1 prostate epithelial cells were less sensitive to ZP and more sensitive to TPEN , but total cellular zinc levels were changed similarly . ZnSO4 enhanced the toxicity of ZP , but inhibited the effects of TPEN as expected . The morphological/biochemical responses to ZP and TPEN differed . ZP decreased ATP levels and stimulated ERK , AKT and PKC phosphorylation . DNA laddering was observed only at low doses of ZP but all doses of TPEN . TPEN activated caspase 3/7 and induced PARP-cleavage , DNA-fragmentation , ROS-formation and apoptotic bodies . PKC and ERK-pathway inhibitors , and antioxidants protected against ZP-induced but not TPEN-induced death . Inhibitors of MPTP-opening protected both . Cell death in response to TPEN ( but not ZP ) was diminished by a calpain inhibitor and largely prevented by a caspase 3 inhibitor . Overall , the results indicated primarily a necrotic cell death for ZP and an apoptotic cell death for TPEN . The enhanced sensitivity of PCa cells to ZP and the apparent ability of ZP and TPEN to kill quiescent and rapidly dividing cells in a p53-independent manner suggest that ZP/TPEN might be used to develop adjunct treatments for PCa . OUTPUT: resisting cell death INPUT: Overexpression of the adverse prognostic marker ERBB2 occurs in 30% of breast cancers ; however , therapies targeting this gene have not proved to be as effective as was initially hoped . Transcriptional profiling meta-analyses have shown that there are approximately 150 genes co-overexpressed with ERBB2 , suggesting that these genes may represent alternative factors influencing ERBB2-positive tumors . Here we describe an RNA interference-based analysis of these genes that identifies transcriptional regulators of fat synthesis and storage as being critical for the survival of these cells . These transcription factors , nuclear receptor subfamily 1 , group D , member 1 ( NR1D1 ) and peroxisome proliferator activated receptor gamma binding protein ( PBP ) , both reside on ERBB2-containing 17q12-21 amplicons and are part of the ERBB2 expression signature . We show that NR1D1 and PBP act through a common pathway in upregulating several genes in the de novo fatty acid synthesis network , which is highly active in ERBB2-positive breast cancer cells . Malate dehydrogenase 1 and malic enzyme 1 , enzymes that link glycolysis and fatty acid synthesis , are also regulated by NR1D1 . The resulting high-level fat production from increased expression of these genes likely contributes to an abnormal cellular energy metabolism based on aerobic glycolysis . Together , these results show that the cells of this aggressive form of breast cancer are genetically preprogrammed to depend on NR1D1 and PBP for the energy production necessary for survival . OUTPUT: cellular energetics INPUT: Bisphenol A ( BPA ) is one of the most prevalent chemicals in daily-use materials , therefore , human exposure to BPA is ubiquitous . We found that low concentrations of BPA stimulate the spermatogonial GC-1 cells proliferation by G protein-coupled receptor 30 ( GPR30)-mediated epidermal growth factor receptor ( EGFR)-extracellular regulated kinase ( ERK)-c-Fos pathway . However , through the same pathway GPR30 expression has been shown to be induced by EGF , an EGFR ligand . Thus , we want to know if low concentrations of BPA are able to induce the GPR30 expression and the possible mechanism(s) in GC-1 cells . By transient transfection with expression plasmids , 10(-9)M BPA significantly transactivates the Gpr30-5'-flanking region through activating the GPR30 , cGMP-dependent protein kinase ( PKG ) , estrogen receptor-α ( ER-α ) , and EFGR-ERK pathways . Furthermore , an activator protein-1 ( AP-1 ) site located within this region is found to be responsible for the transactivation of BPA . Expectedly , through the same pathways , BPA significantly induces the gene and protein expression of GPR30. c-Fos is further observed to be strongly recruited to the AP-1 site in a chromatin immunoprecipitation assay and its dysfunction on the AP-1 site markedly suppresses the expression of GPR30 , p-ERK1/2 , p-Ser118-ER-α and cell proliferation by BPA . Our results demonstrate that a low-concentration BPA induces GPR30 expression through the GPR30-EFGR-ERK-c-Fos , ER-α , and PKG pathways , presumably boosting the cells proliferation via a regulatory loop . The present study provides a novel insight into the potential role of GPR30 in the initiation and progression of male germ cell cancer induced by environmentally relevant BPA . OUTPUT: sustaining proliferative signaling INPUT: Piceatannol has potent anti-inflammatory , immunomodulatory , anticancer and antiproliferative effects . However , little is known about the mechanism by which piceatannol inhibits invasion and metastasis . The aim of the current study was to investigate the effects of piceatannol on the expression of matrix metalloproteinase-9 ( MMP-9 ) in DU145 human prostate cancer cells . The results revealed that MMP-9 activity was significantly increased in response to tumor necrosis factor-α ( TNF-α ) . However , treatment with piceatannol reversed TNF-α- and MMP-9-induced gelatin zymography and its gene expression . In addition , a Matrigel invasion assay determined that piceatannol reduces the TNF-α-induced invasion of DU145 cells . Nuclear factor-κ B ( NF-κB ) is a significant transcription factor that regulates numerous genes involved in tumor cell invasion and metastasis . Therefore , whether piceatannol acts on NF-κB to regulate MMP-9 gene expression was analyzed . The results revealed that piceatannol attenuates MMP-9 gene expression via the suppression of NF-κB activity . Using a specific NF-κB inhibitor , pyrrolidine dithiocarbamate , it was confirmed that TNF-α-induced MMP-9 gene expression is primarily regulated by NF-κB activation . Piceatannol inhibited NF-κB activity by suppressing nuclear translocation of the NF-κB p65 and p50 subunits . Furthermore , TNF-α-induced Akt phosphorylation was significantly downregulated in the presence of piceatannol . The Akt inhibitor LY294002 caused a significant decrease in TNF-α-induced NF-κB activity and MMP-9 gene expression . Overall , these data suggest that piceatannol inhibits TNF-α-induced invasion by suppression of MMP-9 activation via the Akt-mediated NF-κB pathway in DU145 prostate cancer cells . OUTPUT: activating invasion and metastasis INPUT: ZBP-89 ( ZNF148 ) is a Zinc finger Binding Protein of 89 kDa that binds GC-rich DNA elements . Originally , it was expression cloned using a DNA element mediating EGF regulation of the gastrin promoter . ZBP-89 functions as both a transcriptional activator and repressor . A variety of extracellular regulators including TGFbeta , retinoic acid and butyrate stimulate ZBP-89 gene expression . Butyrate activation of p21WAF1 is potentiated by ZBP-89 through the recruitment of the co-activator p300 , while chronic stimulation by butyrate increases ZBP-89 gene expression correlating with cell differentiation . ZBP-89 stimulates growth arrest and apoptosis through its ability to bind the p21WAF1 promoter or its ability to form protein-protein interactions with p53 . ZBP-89 protein is elevated in a variety of gastrointestinal cancers as well as the pancreas . In particular , ZBP-89 is normally expressed in pancreatic islets and ducts and in about 30% of pancreatic adenocarcinomas . OUTPUT:

evading growth suppressors;resisting cell death

HoC_dynamic_5_shot51

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Colorectal cancer ( CRC ) arises as the consequence of progressive changes from normal epithelial cells through polyp to tumor , and thus is an useful model for studying metabolic shift . In the present study , we studied the metabolomic profiles using high analyte specific gas chromatography/mass spectrometry ( GC/MS ) and liquid chromatography tandem mass spectrometry ( LC/MS/MS ) to attain a systems-level view of the shift in metabolism in cells progressing along the path to CRC . Colonic tissues including tumor , polyps and adjacent matched normal mucosa from 26 patients with sporadic CRC from freshly isolated resections were used for this study . The metabolic profiles were obtained using GC/MS and LC/MS/MS . Our data suggest there was a distinct profile change of a wide range of metabolites from mucosa to tumor tissues . Various amino acids and lipids in the polyps and tumors were elevated , suggesting higher energy needs for increased cellular proliferation . In contrast , significant depletion of glucose and inositol in polyps revealed that glycolysis may be critical in early tumorigenesis . In addition , the accumulation of hypoxanthine and xanthine , and the decrease of uric acid concentration , suggest that the purine biosynthesis pathway could have been substituted by the salvage pathway in CRC . Further , there was a step-wise reduction of deoxycholic acid concentration from mucosa to tumors . It appears that to gain a growth advantage , cancer cells may adopt alternate metabolic pathways in tumorigenesis and this flexibility allows them to adapt and thrive in harsh environment . OUTPUT: cellular energetics INPUT: The contribution that mitochondrial bioenergetics could have in cancer development is debated . Here , we have generated HCT116-derived colocarcinoma cell lines expressing different levels of the beta catalytic subunit of the mitochondrial H+-adenosine triphosphate synthase to assess the contribution of mitochondrial bioenergetics in colon cancer progression . The generated cells exhibit large ultrastructural , transcriptomic , proteomic and functional differences in their mitochondria and in their in vivo tumor forming capacity . We show that the activity of oxidative phosphorylation defines the rate of glucose utilization by aerobic glycolysis . The aggressive cellular phenotype , which is highly glycolytic , is bound to the deregulated expression of genes involved in metabolic processes , the regulation of the cell cycle , apoptosis , angiogenesis and cell adhesion . Remarkably , the molecular and ultrastructural analysis of the tumors derived from the three HCT116 cell lines under study highlight that tumor promotion inevitably requires the selection of cancer cells with a repressed biogenesis and functional activity of mitochondria , i.e. the highly glycolytic phenotype is selected for tumor development . The tumor forming potential of the cells is a non-genetically acquired condition that provides the cancer cell with a cell-death resistant phenotype . An abrogated mitochondrial respiration contributes to a diminished potential for reactive oxygen species signaling in response to 5-fluorouracil treatment . Treatment of cancer cells with dichloroacetate partially restores the functional differentiation of mitochondria and promotes tumor regression , emphasizing the reversible nature of the metabolic trait of cancer . OUTPUT: cellular energetics INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Calcium ( Ca(2+) ) signals are involved in important checkpoints in cell death pathways and promote both apoptosis and autophagy . However , the relationship between autophagy and apoptosis in response to Ca(2+) level elevation is poorly understood . Here , we provided evidence that the influx of extracellular Ca(2+) triggered by Trichokonin VI ( TK VI ) , an antimicrobial peptide , induced calpain-dependent apoptosis and autophagy in hepatocellular carcinoma ( HCC ) cells . Remarkably , TK VI preferentially induced apoptosis that was associated with calpain-mediated Bax and Atg5 cleavage , which resulted in the collapse of the mitochondrial membrane potential and cytochrome c release . Interestingly , truncated , but not full-length Atg5 , associated with Bcl-xL and promoted the intrinsic pathway . Moreover , TK VI treatment induced reactive oxygen species ( ROS ) accumulation , an effect in which Bak might play a major role . This accumulation of ROS resulted in the subsequent disposal of damaged mitochondria within autophagosomes via Atg5-mediated and mitochondria-selective autophagy . Both the inhibition of calpain activity and Bax deficiency activated a switch that promoted an enhancement of autophagy . The inhibition of both apoptosis and autophagy significantly attenuated the TK VI cytotoxicity , indicating that the two processes had stimulatory effects during TK VI-meditated cell death . These results suggested that calpain , Bak and Atg5 were molecular links between autophagy and apoptosis and revealed novel aspects of the crosstalk between these two processes . The potential of TK VI is proposed as a promising anticancer agent for its well-characterized activity of Ca(2+) agonist and as a possible novel therapeutic strategy that acts on cancer cell mitochondria . OUTPUT: resisting cell death;tumor promoting inflammation INPUT: Twelve postmenopausal women ( 54-93 years ) with primary breast carcinoma were treated with tamoxifen due to infirmity or refusal to undergo surgery . Seven premenopausal patients ( 32-50 years ) were given preoperative chemotherapy because of large tumors or inflammatory carcinoma . Fine-needle aspiration biopsy was used to procure tumor cells for diagnosis , hormone receptor determination and analysis of proliferation fraction . Aspirations were repeated every 3 months in the tamoxifen group and each month in patients receiving chemotherapy . Two patients who responded to tamoxifen had tumors with more than 75% estrogen receptor positive cells . A decreased proliferation fraction was observed in two tumors responding to tamoxifen . Eight patients , all with estrogen receptor positive tumors , had stable disease . Progressive disease was observed in two patients with less than 25% receptor positive cells . In these tumors the percentage of proliferating cells remained high during therapy . Objective response was recorded for six patients treated with chemotherapy . The clinical response was reflected in a decreased proliferation fraction . No correlation was observed between response and percentage of proliferating cells in the untreated tumor . The results suggest that analysis of tumor cell characteristics such as hormone receptor content and proliferation fraction can be used to predict and monitor response to endocrine treatment and chemotherapy in breast carcinomas . OUTPUT: sustaining proliferative signaling INPUT: The purpose of this study was to use the proteomics approach , which is based on high resolution two-dimensional electrophoresis coupled with multivariate correspondence analysis and mass spectrometry , to classify objectively the biochemical basis of the anti-cancer activity of the synthetic cyclin-dependent kinase inhibitor , bohemine ( BOH ) . The changes in the cell cycle and corresponding protein composition of the A549 human lung adenocarcinoma cell line after treatment with BOH were evaluated and proteins differentially expressed in the BOH treated A549 cells , compared to the untreated A549 counterparts , were selected . Thirteen of these candidate proteins associated with the drug effects in vitro were identified by mass spectrometry . Many of these proteins fall into one of three functional categories : i ) metabolic pathways ( glycolysis , nucleic acid synthesis and NADPH production ) , ii ) stress response and protein folding , and iii ) cytoskeleton and exocytosis . Changes in protein expression patterns corresponded to a higher resistance of A549 lung carcinoma cells to BOH when compared to the CEM leukaemia cell line . These protein changes reflect a fine balance of the resistant versus the susceptible phenotype in response to the drug . Since BOH is a selective cyclin-dependent kinase inhibitor , changes in the protein expression pattern can be more generally associated with cell cycle regulation as evidenced by inhibition of cell cycling in A549 cells . Our conclusions further underline the importance of cell cycle control in both the cellular signalling and metabolic pathways . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot52

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Signal transducers and activators of transcription 3 ( Stat3 ) is activated by cytokines and growth factors in lung cancers and regulates expression of genes implicated in cell growth , survival , and transformation . Previously , we found that mice with a deletion of the G protein-coupled receptor , family C , group 5 , member a ( Gprc5a ) gene develop lung tumors , indicating that Gprc5a is a tumor suppressor . Herein , we show that epithelial cells from Gprc5a knockout mouse lung ( Gprc5a(-/-) cells ) survive better in vitro in medium deprived of exogenous growth factors and form more colonies in semisolid medium than their counterparts from wild-type mice ( Gprc5a(+/+) cells ) . Stat3 tyrosine 705 phosphorylation and expression of several Stat3-regulated antiapoptotic genes were higher in Gprc5a(-/-) than in Gprc5a(+/+) cells . Both cell types secreted leukemia inhibitory factor ( Lif ) ; however , whereas Stat3 activation was persistent in Gprc5a(-/-) cells , it was transient in Gprc5a(+/+) cells . Lung adenocarcinoma cells isolated from Gprc5a(-/-) mice also exhibited autocrine Lif-mediated Stat3 activation . The level of Socs3 , the endogenous Stat3 inhibitory protein , was higher in Gprc5a(+/+) than in Gprc5a(-/-) cells , and expression of the tumor suppressor stabilized Socs3 . Inhibition of Stat3 signaling in Gprc5a(-/-) normal and cancer cells by the Janus-activated kinase 2 inhibitor AG490 or by a dominant negative Stat3(Y705F) increased starvation-induced apoptosis and inhibited colony formation . These results show that persistent Stat3 activation is important for the survival and transformation of Gprc5a(-/-) lung cells and suggest that the tumor suppressive effects of Gprc5a are mediated , at least in part , by inhibition of Stat3 signaling through Socs3 stabilization . OUTPUT: sustaining proliferative signaling INPUT: SET and MYND domain-containing protein 3 ( SMYD3 ) is a histone methyltransferase that plays an important role in transcriptional regulation in human carcinogenesis . It can specifically methylate histone H3 at lysine 4 and activate the transcription of a set of downstream genes , including several oncogenes ( e.g. , N-myc , CrkL , Wnt10b , RIZ and hTERT ) and genes involved in the control of cell cycle ( e.g. , CyclinG1 and CDK2 ) and signal transduction ( e.g. , STAT1 , MAP3K11 and PIK3CB ) . To determine the effects of SMYD3 over-expression on cell proliferation , we transfected SMYD3 into MDA-MB-231 cells and found that these cells showed several transformed phenotypes as demonstrated by colony growth in soft agar . Besides , we show here that down-regulation of SMYD3 could induce G1-phase cell cycle arrest , indicating the potent induction of apoptosis by SMYD3 knockdown . These results suggest the regulatory mechanisms of SMYD3 on the acceleration of cell cycle and facilitate the development of strategies that may inhibit the progression of cell cycle in breast cancer cells . OUTPUT: evading growth suppressors;resisting cell death INPUT: Angiogenesis is critical for cancer growth and metastasis . Steps of angiogenesis are energy consuming , while vascular endothelial cells are highly glycolytic . Glioblastoma multiforme ( GBM ) is a highly vascular tumor and this enhances its aggressiveness . D-amino acid oxidase ( DAO ) is a promising therapeutic protein that induces oxidative stress upon acting on its substrates . Oxidative stress-energy depletion ( OSED ) therapy was recently reported ( El Sayed et al. , Cancer Gene Ther , 19 , 1-18 , 2012 ) . OSED combines DAO-induced oxidative stress with energy depletion caused by glycolytic inhibitors such as 3-bromopyruvate ( 3BP ) , a hexokinase II inhibitor that depleted ATP in cancer cells and induced production of hydrogen peroxide. 3BP disturbs the Warburg effect and antagonizes effects of lactate and pyruvate ( El Sayed et al. , J Bioenerg Biomembr , 44 , 61-79 , 2012 ) . Citrate is a natural organic acid capable of inhibiting glycolysis by targeting phosphofructokinase . Here , we report that DAO , 3BP and citrate significantly inhibited angiogenesis , decreased the number of vascular branching points and shortened the length of vascular tubules . OSED delayed the growth of C6/DAO glioma cells. 3BP combined with citrate delayed the growth of C6 glioma cells and decreased significantly the number and size of C6 glioma colonies in soft agar . Human GBM cells ( U373MG ) were resistant to chemotherapy e.g. cisplatin and cytosine arabinoside , while 3BP was effective in decreasing the viability and disturbing the morphology of U373MG cells . OUTPUT: inducing angiogenesis INPUT: Epigenetic changes in pediatric neuroblastoma may contribute to the aggressive pathophysiology of this disease , but little is known about the basis for such changes . In this study , we examined a role for the DNA methyltransferase DNMT3B , in particular , the truncated isoform DNMT3B7 , which is generated frequently in cancer . To investigate if aberrant DNMT3B transcripts alter DNA methylation , gene expression , and phenotypic character in neuroblastoma , we measured DNMT3B expression in primary tumors . Higher levels of DNMT3B7 were detected in differentiated ganglioneuroblastomas compared to undifferentiated neuroblastomas , suggesting that expression of DNMT3B7 may induce a less aggressive clinical phenotype . To test this hypothesis , we investigated the effects of enforced DNMT3B7 expression in neuroblastoma cells , finding a significant inhibition of cell proliferation in vitro and angiogenesis and tumor growth in vivo . DNMT3B7-positive cells had higher levels of total genomic methylation and a dramatic decrease in expression of the FOS and JUN family members that comprise AP1 transcription factors . Consistent with an established antagonistic relationship between AP1 expression and retinoic acid receptor activity , increased differentiation was seen in the DNMT3B7-expressing neuroblastoma cells following treatment with all-trans retinoic acid ( ATRA ) compared to controls . Our results indicate that DNMT3B7 modifies the epigenome in neuroblastoma cells to induce changes in gene expression , inhibit tumor growth , and increase sensitivity to ATRA . OUTPUT: inducing angiogenesis;sustaining proliferative signaling INPUT: Dimethyl sulfoxide ( DMSO ) , a well-known differentiation inducer in several myeloid cells , also induces a reversible G(1) arrest in many cell lines . We recently showed that DMSO induces a G(1) phase arrest in Chinese hamster ovary ( CHO ) cells , by restoring contact inhibition and preventing high density-dependent apoptosis . CHO cells are frequently used in cell biology and mutagenesis studies due to their good growth capacity and ease of manipulation but are very difficult to synchronize by serum starvation since they detach from monolayers when they reach confluence . In this study we investigated the possibility of using DMSO to reversibly synchronize CHO cells in the G(1) phase of the cell cycle and analysed whether toxic effects follow the arrest using growth curve , sister chromatid exchange and micronuclei assays . We carried out a kinetic analysis of the arrest by DMSO and re-entry into the cell cycle after drug release by cytofluorimetric analysis of DNA content and bromodeoxyuridine incorporation . We show that CHO cells are efficiently and reversibly arrested in G(1) by DMSO in concentrations ranging between 1 and 2% . In our experiments , >90% of cells grown for 96 h in presence of the drug were arrested in G(1) and synchronously re-entered S phase approximately 8-12 h after release . Furthermore , expression levels of p27 were down-regulated during G(1) progression and cyclin D3 and E expression patterns were similar to those observed after serum starvation . No detectable cytotoxicity or genetic damage were induced in G(1) released cells as revealed by the tests employed . Our results show that DMSO is a very powerful inducer of G(1) synchronization in CHO cells without detectable cytotoxic or genetic effects in cell populations released from G(1) arrest . DMSO synchronization represents a model system in which to analyse protein activities regulating G(1) progression and investigate the response of G(1) cells to mutagen treatments . OUTPUT: sustaining proliferative signaling;evading growth suppressors;genomic instability and mutation INPUT: To investigate the significance of sialylation and sulfation of lactosylceramide in transformed cells , we established ganglioside GM3- and lactosylsulfatide ( SM3)-reconstituted cells by transfecting cDNAs of GM3 synthase and cerebroside sulfotransferase into the J5 subclone of 3LL Lewis lung carcinoma cells . The J5 clone was selected for the transfection of these genes because it lacks GM3 and SM3 but accumulates lactosylceramide . The anchorage-dependent growth of both GM3- and SM3-reconstituted cells was similar . However , anchorage-independent growth ( as measured by colony-forming ability in soft agar ) of the SM3- reconstituted cells was almost completely lost , which supports our previous observation showing the suppression of tumorigenic potential in vivo and beta1 integrin gene expression induced by the introduction of cerebroside sulfotransferase gene ( Kabayama et al. [ 2001 ] J. Biol . Chem. , 276 , 26777-26783 ) . The GM3-reconstituted cells formed a significantly higher number of colonies in soft agar compared to mock-transfected cells and began to proliferate and become resistant to apoptosis when serum was depleted , indicating that endogenous GM3 is essential for maintaining these fundamental properties of malignant cells . We also found that serum-induced ERK1/2 activation was suppressed in the GM3-reconstituted cells , suggesting that anchorage-independent cell cycle initiation by endogenous GM3 is elicited through pathway(s) independent of ERK1/2 activation . The selective down-regulation of platelet-derived growth factor ( PDGF)-dependent ERK1/2 activation in the GM3-reconstituted cells was due to the substantial decreases of PDGF alpha receptor mRNA and protein , but in the SM3-reconstituted cells PDGF alpha receptor expression was similar to mock cells . Thus , endogenously produced GM3 and SM3 differentially and distinctly regulate tumor-progression ability , that is , GM3 leads the transformed phenotype of J5 cells to promotion and SM3 to abrogation . OUTPUT:

resisting cell death;sustaining proliferative signaling

HoC_dynamic_5_shot53

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: The identification of molecular markers related to critical biological processes during carcinogenesis may aid in the evaluation of carcinogenic potentials of chemicals and chemical mixtures . Work from our laboratory demonstrated that a single treatment with N-methyl-N'-nitro-N-nitrosoguanidine ( MNNG ) enhanced spontaneous malignant transformation of the human keratinocyte cell line RHEK-1 . In contrast , chronic low-level exposure of cells to arsenic alone or in a mixture containing arsenic , cadmium , chromium , and lead inhibited malignant conversion . To identify changes in gene expression that influence these different outcomes , cDNA microarray technology was used . Analysis of multiple human arrays in MNNG-transformed RHEK-1 cells , designated OM3 , and those treated with arsenic or the arsenic-containing metal mixture showed unique patterns of gene expression . Genes that were overexpressed in OM3 included oncogenes , cell cycle regulators , and those involved in signal transduction , whereas genes for DNA repair enzymes and inhibitors of transformation and metastasis were suppressed . In arsenic-treated cells , multiple DNA repair proteins were overexpressed . Mixture-treated cells showed increased expression of a variety of genes including metallothioneins and integrin 4 . These cells showed decreased expression of oncogenes , DNA repair proteins , and genes involved in the mitogen-activated protein kinase pathway . For comparison we are currently analyzing gene expression changes in RHEK-1 cells transformed by other means . The goal of these studies is to identify common batteries of genes affected by chemical modulators of the carcinogenic process . Mechanistic studies may allow us to correlate alterations in their expression with sequential stages in the carcinogenic process and may aid in the risk assessment of other xenobiotics . OUTPUT: activating invasion and metastasis;evading growth suppressors;genomic instability and mutation;sustaining proliferative signaling INPUT: Although the immense efforts have been made for cancer prevention , early diagnosis , and treatment , cancer morbidity and mortality has not been decreased during last forty years . Especially , lung cancer is top-ranked in cancer-associated human death . Therefore , effective strategy is strongly required for the management of lung cancer . In the present study , we found that novel daphnane diterpenoids , yuanhualine ( YL ) , yuanhuahine ( YH ) and yuanhuagine ( YG ) isolated from the flower of Daphne genkwa ( Thymelaeaceae ) , exhibited potent anti-proliferative activities against human lung A549 cells with the IC50 values of 7.0 , 15.2 and 24.7 nM , respectively . Flow cytometric analysis revealed that the daphnane diterpenoids induced cell-cycle arrest in the G0/G1 as well as G2/M phase in A549 cells . The cell-cycle arrests were well correlated with the expression of checkpoint proteins including the up-regulation of cyclin-dependent kinase inhibitor p21 and p53 and down-regulation of cyclin A , cyclin B1 , cyclin E , cyclin dependent kinase 4 , cdc2 , phosphorylation of Rb and cMyc expression . In the analysis of signal transduction molecules , the daphnane diterpenoids suppressed the activation of Akt , STAT3 and Src in human lung cancer cells . The daphnane diterpenoids also exerted the potent anti-proliferative activity against anticancer-drug resistant cancer cells including gemcitabine-resistant A549 , gefitinib- , erlotinib-resistant H292 cells . Synergistic effects in the growth inhibition were also observed when yuanhualine was combined with gemcitabine , gefitinib or erlotinib in A549 cells . Taken together , these findings suggest that the novel daphnane diterpenoids might provide lead candidates for the development of therapeutic agents for human lung cancers . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Progression to malignancy requires that cells overcome senescence and switch to an immortal phenotype . Thus , exploring the genetic and epigenetic changes that occur during senescence/immortalization may help elucidate crucial events that lead to cell transformation . In the present study , we have globally profiled DNA methylation in relation to gene expression in primary , senescent and immortalized mouse embryonic fibroblasts . Using a high-resolution genome-wide mapping technique , followed by extensive locus-specific validation assays , we have identified 24 CpG islands that display significantly higher levels of CpG methylation in immortalized cell lines as compared to primary murine fibroblasts . Several of these hypermethylated CpG islands are associated with genes involved in the MEK-ERK pathway , one of the most frequently disrupted pathways in cancer . Approximately half of the hypermethylated targets are developmental regulators , and bind to the repressive Polycomb group ( PcG ) proteins , often in the context of bivalent chromatin in mouse embryonic stem cells . Because PcG-associated aberrant DNA methylation is a hallmark of several human malignancies , our methylation data suggest that epigenetic reprogramming of pluripotency genes may initiate cell immortalization . Consistent with methylome alterations , global gene expression analysis reveals that the vast majority of genes dysregulated during cell immortalization belongs to gene families that converge into the MEK-ERK pathway . Additionally , several dysregulated members of the MAP kinase network show concomitant hypermethylation of CpG islands . Unlocking alternative epigenetic routes for cell immortalization will be paramount for understanding crucial events leading to cell transformation . Unlike genetic alterations , epigenetic changes are reversible events , and as such , can be amenable to pharmacological interventions , which makes them appealing targets for cancer therapy when genetic approaches prove inadequate . OUTPUT: enabling replicative immortality INPUT: AIM The study was designed to explore the effects of antisense human telomerase RNA ( ahTR ) on the malignant phenotype of gastric carcinoma cell line SGC-7901 , and its potential role in gene therapy for tumors . METHODS An ahTR eukaryotic expression vector , including the sequence of template region of telomere repeats , was constructed by recombinant technology of molecules and then transfected into gastric carcinoma cell line SGC-7901 by liposome DOTAP . Subsequently , the expression of hTR RNA and ahTR RNA by reverse transcription-polymerase chain reaction , telomerase activity by telomeric repeat amplification protocol-ELISA ( TRAP-ELISA ) , telomere length by Southern blotting , cell morphology under light microscope , cellular proliferation capacity by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay , cell-cycle distribution by flow cytometry , efficiency of clone formation in soft agar , and tumorigenecity in nude mice were examined and evaluated in ahTR-transfected cells , control plasmid pCI-neo transfected cells and their parental cells . RESULTS An ahTR eukaryotic expression vector was constructed and successfully transfected into SGC-7901 cells . The telomerase activity in ahTR-transfected SGC-7901 cells decreased from 100% to approximately 25% , and telomere length in the cells shortened to 3.35 from 4.08 Kb at 60 population doublings . Compared with the parental cells and pCI-neo transfected cells , ahTR-transfected cells displayed some morphological changes , such as decreased atypia , and recovery of contact inhibition and density inhibition under light microscope . Furthermore , ahTR-transfected cells displayed decreased invasive capacity in Borden's chamber invasive model , increased G0/G1 phase rate and apoptotic rate . Surprisingly , ahTR-transfected SGC-7901 cells lost their capacity for clone formation in soft agar and tumorigencity in nude mice . CONCLUSION Antisense-hTR transfection can inhibit the growth of SGC-7901 cells and partially reverse the malignant phenotypes . This study provides an exciting approach for cancer therapy by inhibiting telomerase activity using an antisense gene . OUTPUT: evading growth suppressors;activating invasion and metastasis;resisting cell death INPUT: Identification of the proteins that are associated with estrogen receptor ( ER ) status is a first step towards better understanding of the hormone-dependent nature of breast carcinogenesis . Although a number of gene expression analyses have been conducted , protein complement has not been systematically investigated to date . Because proteins are primary targets of therapeutic drugs , in this study , we have attempted to identify proteomic signatures that demarcate ER-positive and -negative breast cancers . Using highly enriched breast tumor cells , replicate analyses from 3 ERα+ and 3 ERα- human breast tumors resulted in the identification of 2,995 unique proteins with ≥2 peptides . Among these , a number of receptor tyrosine kinases and intracellular kinases that are abundantly expressed in ERα+ and ERα- breast cancer tissues were identified . Further , label-free quantitative proteome analysis revealed that 236 proteins were differentially expressed in ERα+ and ERα- breast tumors . Among these , 141 proteins were selectively up-regulated in ERα+ , and 95 proteins were selectively up-regulated in ERα- breast tumors . Comparison of differentially expressed proteins with a breast cancer database revealed 98 among these have been previously reported to be involved in breast cancer . By Gene Ontology molecular function , dehydrogenase , reductase , cytoskeletal proteins , extracellular matrix , hydrolase , and lyase categories were significantly enriched in ERα+ , whereas selected calcium-binding protein , membrane traffic protein , and cytoskeletal protein were enriched in ERα- breast tumors . Biological process and pathway analysis revealed that up-regulated proteins of ERα+ were overrepresented by proteins involved in amino acid metabolism , proteasome , and fatty acid metabolism , while up-regulated proteins of ERα- were overrepresented by proteins involved in glycolysis pathway . The presence and relative abundance of 4 selected differentially abundant proteins ( liprin-α1 , fascin , DAP5 , and β-arrestin-1 ) were quantified and validated by immunohistochemistry . In conclusion , unlike in vitro cell culture models , the in vivo signaling proteins and pathways that we have identified directly from human breast cancer tissues may serve as relevant therapeutic targets for the pharmacological intervention of breast cancer . OUTPUT: cellular energetics INPUT: The early stages of head and neck cancer are presumed to require a senes of genetic alterations that are not represented by a distinct clinical phenotype . Therefore , genes with altered expression in the preneoplasia may be useful for the early detection of this highly recurrent cancer . In this study , we immortalized normal human oral keratinocytes ( NHOK ) by retroviral-mediated infection of HPV 16 transforming oncogenes , E6 and E7 ( HOK16E6E7 ) . Using the Affymetrix gene chip ( U95Av2 ) , we identified 177 known genes and EST that were overexpressed at least 3-fold or above in the immortalized cells , while 133 were down-regulated compared to NHOK . Northern blot analysis showed elevated levels of p55CDC in the immortalized cells , while NHOK showed high basal expression of small proline rich protein ( SPRR2 ) . The altered expression of these genes maybe associated with cellular proliferation or differentiation and the early stages of oral carcinogenesis . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot54

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Epidermal growth factor ( EGF ) receptor is inversely related to expression of estrogen receptor ( ER ) and progesterone receptor in primary breast tumors and is a negative predictor for response to endocrine therapy . To investigate a possible causal role of EGF receptor expression in breast cancer progression to hormone independence , we have created an experimental cell system . Epidermal growth factor receptor complementary DNA was introduced in estrogen-dependent ZR-75-1 breast cancer cells , and the resulting ZR/HERc cells exhibited a mitogenic response to epidermal growth factor , thus bypassing estrogen dependence . This EGF-induced proliferation could not be inhibited by antiestrogens . In addition , we noted changes in cell morphology and keratin expression of EGF-stimulated ZR/HERc cells , suggestive of an altered differentiation state . Furthermore , intolerance of functional ER and EGF receptor signal transduction pathways in ZR/HERc cells was observed during simultaneous activation , which possibly explains the inverse relationship of ER and EGF receptor expression in primary tumors . In contrast to the parental cells , ZR/HERc cells rapidly progressed to a stable ER-negative phenotype when cultured in the presence of the antiestrogen hydroxy-tamoxifen . These results suggest a possible role for EGF receptor in progression of breast cancer to hormone independence . OUTPUT: sustaining proliferative signaling INPUT: Findings of increased numbers of epidermal growth factor receptors ( EGF-R ) and increased expression of transforming growth factor alpha ( TGF-alpha ) in surgical specimens of human renal cell carcinoma have led to the proposal that growth of these tumors may be regulated by TGF-alpha in an autocrine manner . In the studies presented here , we have examined this hypothesis using two human renal carcinoma cell lines , SKRC-4 and SKRC-29 . We demonstrated that both SKRC-4 and SKRC-29 cells were growth stimulated by greater than 35% when cultured in the presence of TGF-alpha or EGF and were inhibited by 29% to 46% if cultured in the presence of anti-EGF-R monoclonal antibody 225 . Treatment of cells with TGF-alpha enhanced the levels of expression of EGF-R mRNA and TGF-alpha mRNA . In addition , incubation of cells with monoclonal antibody 225 significantly elevated the levels of excreted TGF-alpha species in the culture medium . Our findings suggest that proliferation of human renal carcinoma cells may be regulated by endogenously produced TGF-alpha and that this regulatory pathway can be interrupted using antibody to its receptor , EGF-R . OUTPUT: sustaining proliferative signaling INPUT: Recent studies indicate that cyclooxygenase-2 ( COX-2 ) is overexpressed in pancreatic adenocarcinoma and may play a critical role in this rapidly progressing form of cancer . A human pancreatic adenocarcinoma cell line , Mia PaCa-2 , was incubated for 18 hours with 5 micromol/L of rofecoxib ( Vioxx ) , a selective COX-2 inhibitor . Total RNA was isolated and gene expression analyzed by DNA microarray chips . In a separate experiment , athymic mice were orthotopically injected with 7.5 x 10(5) Mia PaCa-2 cells through a minilaparotomy . After 1 month , laparotomy was repeated to measure tumor size , and mice were randomized to receive reformulated rodent chow containing either 12.5 mg/kg/day of rofecoxib or no drug for 21 days . Tumor growth was assessed by comparing volume before and after treatment . In vitro , rofecoxib decreased gene expression of cyclin D1/PRAD1 , a key component of cell cycle progression , while increasing expression of several cell cycle arrest genes , including p21/WAF1 , p33/ING , GADD34 , and GADD45 ( P &lt ; 0.05 ) . In vivo , tumor growth was significantly reduced in treated vs. control mice ( P &lt ; 0.05 ) . No systemic toxicity was observed in mice receiving rofecoxib . These data suggest that rofecoxib slows the growth of human pancreatic cancer through changes in gene expression that favor cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Gefitinib , the specific inhibitor of the epidermal growth factor receptor ( EGFR ) , may cause growth delay in cancer cell lines . Thorough understanding of the downstream cellular signaling of gefitinib will facilitate the discovery of biomarkers for predicting outcomes and monitoring anti-EGFR therapies , and provide information for key targets for therapeutic intervention . In this study , we investigated the role of transducer of erbB2.1 ( TOB1 ) in gefitinib therapy . Using the lung carcinoma cell lines A549 and NCI-H1975 , the results suggested that gefitinib might mediate cell cycle arrest in lung cancer cells at least by targeting TOB1 expression . Gefitinib treatment caused cell cycle arrest predominantly at the G1 phase , which is associated with TOB1 nuclear translocation and its interaction with cyclin D1 . We also showed that knockdown of TOB1 expression by RNAi rescued lung cancer cells from gefitinib-induced cell-proliferative arrest . These results suggest that TOB1 interaction with cyclin D1 and nuclear translocation is directly involved in the gefitinib-induced anti-proliferative cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Epidermal growth factor receptor-tyrosine kinase inhibitors ( EGFR-TKIs ) show dramatic antitumor activity in a subset of patients with non-small cell lung cancer who have an active mutation in the epidermal growth factor receptor ( EGFR ) gene . On the other hand , some lung cancer patients with wild type EGFR also respond to EGFR-TKIs , suggesting that EGFR-TKIs have an effect on host cells as well as tumor cells . However , the effect of EGFR-TKIs on host microenvironments is largely unknown . A multiple organ metastasis model was previously established in natural killer cell-depleted severe combined immunodeficient mice using human lung cancer cells . This model was used to investigate the therapeutic efficacy of erlotinib , an EGFR-TKI , on multiple organ metastases induced by human small cell lung cancer cells ( SBC-5 cells ) that did not express EGFR . Although erlotinib did not have any effect on the proliferation of SBC-5 cells in vitro , it significantly suppressed bone and lung metastases in vivo , but not liver metastases . An immunohistochemical analysis revealed that , erlotinib significantly suppressed the number of osteoclasts in bone metastases , whereas no difference was seen in microvessel density . Moreover , erlotinib inhibited EGF-induced receptor activator of nuclear factor kappa-B expression in an osteoblastic cell line ( MC3T3-E1 cells ) . These results strongly suggested that erlotinib prevented bone metastases by affecting host microenvironments irrespective of its direct effect on tumor cells . OUTPUT: activating invasion and metastasis;inducing angiogenesis;sustaining proliferative signaling INPUT: Recent epidemiological and experimental investigations suggest a close relationship between cyclooxygenase ( COX ) and pathogenesis of colorectal cancer . There are two isoforms , COX-1 and COX-2 , which differ in physiological functions and distribution . This study is to investigate the possible roles of both isoforms in the proliferation of colon carcinoma cells . A human colon carcinoma cell line , COLO 320DM , was transfected with an eukaryotic expression vector ( pEF-BOS ) carrying cDNA of either COX-1 or COX-2 . Both COX-1 and COX-2-expressing cells exhibited a similar enzyme activity , 8-10 nmol/10 min/mg of protein . Growth rates of both COX-expressing cells were increased by about 2 fold as compared with mock-transfected cells . The stimulated growth of the COX-expressing cells was confirmed by the increased DNA synthesis as assessed by [ 3H]thymidine incorporation . Furthermore , expression of epidermal growth factor receptor ( EGFR ) was markedly increased in the COX-expressing cells as examined by reverse transcriptase-polymerase chain reaction ( RT-PCR ) . A COX inhibitor , indomethacin , suppressed the stimulated growth , increased DNA synthesis and induction of epidermal growth factor receptor in the COX-1 and COX-2-transfected cells . These results suggest that not only COX-2 but COX-1 is involved in the proliferation of human colon carcinoma cells through the induction of EGFR . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot55

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: The cellular mechanisms of anti-androgen-induced tumor regression have not been investigated in great detail . We have compared the induction of cell death in the androgen-dependent , non-invasive LNCaP prostate cancer cell line by Casodex and TNF-alpha . Both agents induce a dose and time-dependent decrease in cell viability in vitro . However , Casodex does not induce classical DNA fragmentation to oligonucleosomes typically induced by TNF-alpha , but rather induces cleavage to form intermediate 60 kb DNA fragments . RT-PCR based analysis demonstrates that in LNCaP cells Casodex coordinately alters the expression of steady-state level of mRNAs of several matrix metalloproteases and their cognate inhibitors ( most notably MMP2 and TIMP1 ) . Zymography and reverse zymography confirm that the ratio of metalloprotease(s) to inhibitor(s) is altered in favor of activation of the proteases . In a small percentage of the treated LNCaP cells , the activation of the extracellular matrix ( ECM)-proteases by Casodex also induces an invasive phenotype . The acquisition of an invasive phenotype is not seen when LNCaP cells are treated with TNF-alpha , and is not seen when the LNCaP cells are treated with both compounds simultaneously , suggesting that the phenomenon may be specific to particular classes of compounds . These observations have significant implications in the treatment of prostate cancer , since the appearance of a more aggressive phenotype following treatment is clearly undesirable . OUTPUT: resisting cell death;activating invasion and metastasis INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: The immune system has an important role in tumor appearance and spreading . One of the most efficient subpopulations of cytotoxic cells in the destruction of tumors are NK cells . NK cells are activated and increase their cytotoxic potential and modulate their cytokine production after treatment with IFNgamma , IL-12 , TNFalpha and IL-2 . The investigation of the activity of NK cells was performed on peripheral blood lymphocytes ( PBL ) of 16 healthy controls and of 40 patients with metastatic breast carcinoma . Modulation of NK cells was performed with IL-2 , IL-7 , IL-12 , TNFalpha , monoclonal antibodies ( mAb ) for TNFalpha and TNFalpha receptors type I and II , as well as with sera of healthy controls and patients with breast cancer in different clinical stages . Modulating effect of the applied factors after in vitro treatment of PBL was evaluated by the cytotoxic assay using 51chromium . Our results indicate that IL-2 significantly increased the activity of NK cells of controls and breast cancer patients . The sera of patients with advanced breast cancer significantly reduced NK cell activity . IL-7 , IL-12 and mAb for TNFalpha do not significantly change the activity of NK cells . The presence of anti-TNFalpha mAb did not change the inhibitory effect of the sera of breast cancer patients with advanced disease on the activity of NK cells of controls and patients with breast cancer . Blocking of TNFalpha Rcs with mAbs decrease the reactivity of NK cells for IL-2 . The treatment of breast cancer patients with advanced clinical stage of breast cancer with IL-2 , as an additional therapy , could be advantageous , as NK cells after this treatment increase their cytotoxic activity against tumor cells and can improve therapeutical results . OUTPUT: avoiding immune destruction INPUT: INTRODUCTION Necrosis at the tumor center is a common feature of aggressive breast cancers and has been associated with poor prognosis . It is commonly identified by means of invasive histopathology , which often correlates with morbidity and potential tumor cell dissemination , and limits the reconstruction of the whole necrotic domain . In this study we hypothesized that non covalent association to serum albumin ( SA ) and covalent binding to ligands for tumor-abundant cell receptors should synergistically drive selective accumulation and prolonged retention of imaging and therapeutic agents in breast tumor necrotic domains enabling in vivo identification , imaging and possibly treatment of such tumors . METHODS Cyclo-Arg-Gly-Asp-D-Phe-Lys ( c(RGDfK) ) were conjugated to bacteriochlorophyll-derivatives ( Bchl-Ds ) , previously developed as photodynamic agents , fluorescent probes and metal chelators in our lab . The c(RGDfK) component drives ligation to alphaVbeta3 integrin receptors over-expressed by tumor cells and neo-vessels , and the Bchl-D component associates to SA in a non-covalent manner . STL-6014 , a c(RGDfK)-Bchl-D representative , was i.v. injected to CD-1 , nude female mice bearing necrotic and non-necrotic human MDA-MB-231-RFP breast cancer tumors . The fluorescence signals of the Bchl-Ds and RFP were monitored over days after treatment , by quantitative whole body imaging and excised tumor/tissue samples derived thereof . Complementary experiments included competitive inhibition of STL-6014 uptake by free c(RGDfK) , comparative pharmacokinetics of nonconjugated c(RGDfK) Bchl-D ( STL-7012 ) and of two human serum albumin ( HSA ) conjugates : HSA-STL-7012 and HSA-STL-6014 . RESULTS STL-6014 and STL-7012 formed complexes with HSA ( HSA/STL-6014 , HSA/STL-7012 ) . STL-6014 , HSA-STL-7012 and HSA-STL-6014 , selectively accumulated at similar rates , in tumor viable regions over the first 8 h post administration . They then migrated into the necrotic tumor domain and presented tumor half lifetimes ( T1/2 ) in the range of days where T1/2 for HSA-STL-6014 &gt ; STL-6014 &gt ; HSA-STL-7012 . No accumulation of STL-7012 was observed . Pre-injection of c(RGDfK) excess , prevented the uptake of STL-6014 in the small , but not in the large tumors . CONCLUSIONS Non-covalent association to SA and covalent binding to c(RGDfK) , synergistically enable the accumulation and prolonged retention of Bchl-Ds in the necrotic regions of tumors . These findings provide novel guidelines and strategy for imaging and treatment of necrotic tumors . OUTPUT: resisting cell death INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Neutrophils have become recognised as important contributors to the effectiveness of tumour eradication by photodynamic therapy ( PDT ) . In this study , we have used the mouse SCCVII squamous cell carcinoma model to investigate the activity of neutrophils in tumours treated by PDT . Tumour levels of neutrophilic myeloperoxidase ( MPO ) demonstrated not only a massive and sustained sequestration of these cells in PDT-treated tumours but also revealed their activated state evidenced by the presence of released MPO . Among the adhesion molecules expressed on tumour vascular endothelium , ICAM-1 appears to be of primary importance in the invasion of neutrophils into PDT-treated tumours , because its functional blocking with monoclonal antibodies reduced the tumour cure rate . A marked upregulation of its ligands CD11b/CD18 and CD11c/CD18 found on neutrophils associated with PDT-treated tumours supports this assumption . To evaluate the role of inflammatory cytokines regulating neutrophil activity , neutralising antibodies were given to mice before PDT treatment . The results suggest that IL-1beta activity is critical for the therapeutic outcome , since its neutralisation diminished the cure rates of PDT-treated tumours . No significant effect was observed with anti-IL-6 and anti-TNF-alpha treatment . Further flow cytometry-based examination of neutrophils round in PDT-treated tumours revealed that these cells express MHC class II molecules , which suggests their engagement as antigen-presenting cells and involvement in the development of antitumour immune response . OUTPUT:

tumor promoting inflammation

HoC_dynamic_5_shot56

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: γ-Tocotrienol and sesamin are phytochemicals that display potent anticancer activity . Since sesamin inhibits the metabolic degradation of tocotrienols , studies were conducted to determine if combined treatment with sesamin potentiates the antiproliferative effects of γ-tocotrienol on neoplastic mouse ( +SA ) and human ( MCF-7 and MDA-MB-231 ) mammary cancer cells . Results showed that treatment with γ-tocotrienol or sesamin alone induced a significant dose-responsive growth inhibition , whereas combination treatment with these agents synergistically inhibited the growth of +SA , MCF-7 and MDA-MB-231 mammary cancer cells , while similar treatment doses were found to have little or no effect on normal ( mouse CL-S1 and human MCF-10A ) mammary epithelial cell growth or viability . However , sesamin synergistic enhancement of γ-tocotrienol-induced anticancer effects was not found to be mediated from a reduction in γ-tocotrienol metabolism . Rather , combined treatment with subeffective doses of γ-tocotrienol and sesamin was found to induce G1 cell cycle arrest , and a corresponding decrease in cyclin D1 , CDK2 , CDK4 , CDK6 , phospho-Rb , and E2F1 levels , and increase in p27 and p16 levels . Additional studies showed that the antiproliferative effect of combination treatment did not initiate apoptosis or result in a decrease in mammary cancer cell viability . Taken together , these findings indicate that the synergistic antiproliferative action of combined γ-tocotrienol and sesamin treatment in mouse and human mammary cancer cells is cytostatic , not cytotoxic , and results from G1 cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Helicobacter hepaticus-infected Rag2(-/-) mice emulate many aspects of human inflammatory bowel disease , including the development of colitis and colon cancer . To elucidate mechanisms of inflammation-induced carcinogenesis , we undertook a comprehensive analysis of histopathology , molecular damage , and gene expression changes during disease progression in these mice . Infected mice developed severe colitis and hepatitis by 10wk post-infection , progressing into colon carcinoma by 20wk post-infection , with pronounced pathology in the cecum and proximal colon marked by infiltration of neutrophils and macrophages . Transcriptional profiling revealed decreased expression of DNA repair and oxidative stress response genes in colon , but not in liver . Mass spectrometric analysis revealed higher levels of DNA and RNA damage products in liver compared to colon and infection-induced increases in 5-chlorocytosine in DNA and RNA and hypoxanthine in DNA . Paradoxically , infection was associated with decreased levels of DNA etheno adducts . Levels of nucleic acid damage from the same chemical class were strongly correlated in both liver and colon . The results support a model of inflammation-mediated carcinogenesis involving infiltration of phagocytes and generation of reactive species that cause local molecular damage leading to cell dysfunction , mutation , and cell death . There are strong correlations among histopathology , phagocyte infiltration , and damage chemistry that suggest a major role for neutrophils in inflammation-associated cancer progression . Further , paradoxical changes in nucleic acid damage were observed in tissue- and chemistry-specific patterns . The results also reveal features of cell stress response that point to microbial pathophysiology and mechanisms of cell senescence as important mechanistic links to cancer . OUTPUT: tumor promoting inflammation;genomic instability and mutation;resisting cell death INPUT: Investigation has been conducted to delineate the action of some phenolic compounds of natural origin in four human tumor cell lines : acute myeloblastic leukemia ( HL-60 ) , chronic myelogenic leukemia ( K-562 ) , breast adenocarcinoma ( MCF-7 ) and cervical epithelial carcinoma ( HeLa ) . In cells grown in appropriate media the phenolics curcumin , yakuchinone B , resveratrol and capsaicin exhibited growth inhibition as assessed by trypan blue dye exclusion . It was evident from the results of the MTT reduction assay and [ (3)H]thymidine incorporation into nuclear DNA that the phenolics were cytotoxic and inhibited cell proliferation . Dose response studies indicated curcumin to be most cytotoxic towards HL-60 , K-562 and MCF-7 but did not show much activity in HeLa cells . On the other hand , yakuchinone B , although less active than curcumin , displayed cytotoxicity towards all four cell lines . Resveratrol was cytotoxic only in leukemic cells , while capsaicin was marginally cytotoxic . All these phenolics did not elicit any cytotoxic activity as judged by the above parameters towards lymphocytes purified from normal human blood . When cells treated with phenolics were stained with propidium iodide and examined under a fluorescent microscope , characteristic apoptotic features such as chromatin condensation and nuclear fragmentation were observed . Scoring of cells with apoptotic and non-apoptotic features showed positive correlation of apoptotic index with dose of phenolic , and fragmented DNA extracted free of genomic DNA displayed on gel electrophoresis a typical ladder pattern . These phenolics which have human exposure are known cancer chemopreventive agents and their action as inducers of apoptosis in tumor cells suggest their potential use in a strategy for cancer control . OUTPUT:

resisting cell death

HoC_dynamic_5_shot57

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Dietary antioxidants , such as the carotenoids , may protect DNA from oxidative damage . This has been proposed to explain the epidemiological association between higher consumption of fruits and vegetables , which are rich in antioxidants , and lower incidence of cancer . However , this remains to be demonstrated conclusively . The effects of carotenoid supplementation on 1 ) baseline DNA damage , 2 ) susceptibility of cellular DNA to oxidative attack , and 3 ) DNA repair were measured in the human lymphocyte cell line Molt-17 . Baseline DNA damage , susceptibility to oxidant attack ( 100 mumol/l H2O2 for 5 min at 4 degrees C ) , and disappearance of DNA single-strand breaks ( SSB ) after oxidative challenge were monitored by single-cell gel electrophoresis . DNA repair patch synthesis activity in cell extracts was determined using assays that measure nucleotide incorporation during repair of oxidative lesions in template DNA . Unlike single-cell gel electrophoresis , the parameters measured with these assays are not dependent on strand break religation . There was no evidence that beta-carotene , lutein , or beta-cryptoxanthin supplementation protected cellular DNA from oxidation under basal conditions or after oxidative challenge . However , only carotenoid-supplemented cells exhibited a significant decrease in numbers of SSB over a 2-h period after treatment with H2O2 . Carotenoid supplementation did not provoke any detectable change in repair patch synthesis activity . We conclude that supplementation with carotenoids at 8 mumol/l does not provide significant antioxidant protection for DNA in Molt-17 lymphocytes but may enhance recovery of cells from oxidative challenge , as measured by loss of SSB . We argue that these data are most consistent with carotenoids acting to enhance DNA strand break repair . OUTPUT: genomic instability and mutation;tumor promoting inflammation INPUT: UNLABELLED Oxygen free radicals and their reactive derivatives participate in formation of chronic inflammation states , which facilitate development of gastrointestinal tract tumors . Oxidative stress is one of the main causes of damage to cell membranes in result of exacerbated lipid peroxidation process . End products of lipid peroxidation ( aldehydes , organic peroxides ) react with important biological macromolecules such as DNA and proteins , cause changes in cell membrane structure and properties leading to loss of its integrity . Intensification of the lipid peroxidation process is a factor which may also lead to a malfunction in the antioxidant barrier , which further weakens the defense of cells against oxygen free radicals and promotes the onset and development of cancer . The aim of the study was the determination of lipid peroxidation level in gastrointestinal tract tumors ( stomach , liver , colon , and colorectal cancer to liver metastases ) . MATERIAL AND METHODS Materials for studies were obtained from 150 patients with gastrointestinal tract tumors : 10 with stomach cancer , 30 with malignant and benign liver cancers , 60 with primary colorectal cancer , and 50 with metachronous colorectal cancer liver metastases . We also investigated 25 patients with liver cirrhosis , which was treated as a pre-cancerous condition . In total , 175 patients were examined . Tumor specimens , and normal adjacent tissues ( 6-7 cm from the edge of the tumor ) , which served as control tissue in studies , were collected from patients ( with their consent ) during surgery . Additionally , liver specimens were collected from patients with liver cirrhosis . Lipid peroxidation level was determined spectrophotometrically as a concentration of final lipid peroxidation products , which in reaction with tiobarbituric acid ( TBA ) form colour complex ( thiobarbituric acid-reactive substances - TBARS ) . RESULTS The study showed the highest concentration of TBARS in benign , and the lowest in malignant liver tumors . Other types of gastrointestinal tumors studied , were characterized by similar levels of lipid peroxidation . TBARS concentration in these tumors was approximately 2-fold higher than in malignant liver tumors and much lower than in benign tumors . In all cancers of the digestive tract with the exception of malignant liver tumors increased level of TBARS was found , comparing with control tissue . The concentration of TBARS in cirrhotic liver was lower than in control . The level of lipid peroxidation in liver cirrhosis and malignant liver tumors was similar . There were no significant differences in TBARS concentration in the tumors of particular sections of the intestine and normal colon . The highest concentration of TBARS was found in G1 grade of colorectal cancer . In subsequent grades of cells differentiation ( G2 and G3 ) its concentration was lower . The highest level of lipid peroxidation , expressed as the concentration of TBARS was found in the I stage of colorectal cancer clinical advancement . The significantly lowest concentration of TBARS was shown for stage II ( UICC ) . CONCLUSIONS The level of lipid peroxidation in cancerous cells of gastrointestinal tract indicates increased oxidative stress . The changes of lipid peroxidation level--a marker of oxidative stress in gastrointestinal tumors appear to be closely associated with their development stages ( liver cirrhosis/malignant liver cancer ; colorectal cancer/colorectal cancer liver metastases ) and are likely to create such conditions , in which cancerous cells may proliferate , undergo gradual dedifferentiation and malignancy , and generate metastases . OUTPUT: tumor promoting inflammation INPUT: Furan , a potent rodent liver carcinogen , is found in many cooked food items and thus represents a human cancer risk . Mechanisms for furan carcinogenicity were investigated in male F344 rats using the in vivo Comet and micronucleus assays , combined with analysis of histopathological and gene expression changes . In addition , formamidopyrimidine DNA glycosylase ( Fpg ) and endonuclease III ( EndoIII)-sensitive DNA damage was monitored as a measure of oxidative DNA damage . Rats were treated by gavage on four consecutive days with 2 , 4 , and 8mg/kg bw furan , doses that were tumorigenic in 2-year cancer bioassays , and with two higher doses , 12 and 16mg/kg . Rats were killed 3h after the last dose , a time established as producing maximum levels of DNA damage in livers of furan-treated rats . Liver Comet assays indicated that both DNA strand breaks and oxidized purines and pyrimidines increased in a near-linear dose-responsive fashion , with statistically significant increases detected at cancer bioassay doses . No DNA damage was detected in bone marrow , a non-target tissue for cancer , and peripheral blood micronucleus assays were negative . Histopathological evaluation of liver from furan-exposed animals produced evidence of inflammation , single-cell necrosis , apoptosis , and cell proliferation . In addition , genes related to apoptosis , cell-cycle checkpoints , and DNA-repair were expressed at a slightly lower level in the furan-treated livers . Although a mixed mode of action involving direct DNA binding cannot be ruled out , the data suggest that furan induces cancer in rat livers mainly through a secondary genotoxic mechanism involving oxidative stress , accompanied by inflammation , cell proliferation , and toxicity . OUTPUT: genomic instability and mutation;resisting cell death;tumor promoting inflammation;evading growth suppressors INPUT: BACKGROUND Astaxanthin modulates immune response , inhibits cancer cell growth , reduces bacterial load and gastric inflammation , and protects against UVA-induced oxidative stress in in vitro and rodent models . Similar clinical studies in humans are unavailable . Our objective is to study the action of dietary astaxanthin in modulating immune response , oxidative status and inflammation in young healthy adult female human subjects . METHODS Participants ( averaged 21.5 yr ) received 0 , 2 , or 8 mg astaxanthin ( n = 14/diet ) daily for 8 wk in a randomized double-blind , placebo-controlled study . Immune response was assessed on wk 0 , 4 and 8 , and tuberculin test performed on wk 8 . RESULTS Plasma astaxanthin increased ( P &lt ; 0.01 ) dose-dependently after 4 or 8 wk of supplementation . Astaxanthin decreased a DNA damage biomarker after 4 wk but did not affect lipid peroxidation . Plasma C-reactive protein concentration was lower ( P &lt ; 0.05 ) on wk 8 in subjects given 2 mg astaxanthin . Dietary astaxanthin stimulated mitogen-induced lymphoproliferation , increased natural killer cell cytotoxic activity , and increased total T and B cell subpopulations , but did not influence populations of Thelper , Tcytotoxic or natural killer cells . A higher percentage of leukocytes expressed the LFA-1 marker in subjects given 2 mg astaxanthin on wk 8 . Subjects fed 2 mg astaxanthin had a higher tuberculin response than unsupplemented subjects . There was no difference in TNF and IL-2 concentrations , but plasma IFN-gamma and IL-6 increased on wk 8 in subjects given 8 mg astaxanthin . CONCLUSION Therefore , dietary astaxanthin decreases a DNA damage biomarker and acute phase protein , and enhances immune response in young healthy females . OUTPUT: genomic instability and mutation;tumor promoting inflammation INPUT: Breast cancer is the malignant neoplasia with the highest incidence in women worldwide . Chronic oxidative stress and inflammation have been indicated as major mediators during carcinogenesis and cancer progression . Human studies have not considered the complexity of tumor biology during the stages of cancer advance , limiting their clinical application . The purpose of this study was to characterize systemic oxidative stress and immune response parameters in early ( ED ; TNM I and II ) and advanced disease ( AD ; TNM III and IV ) of patients diagnosed with infiltrative ductal carcinoma breast cancer . Oxidative stress parameters were evaluated by plasmatic lipoperoxidation , carbonyl content , thiobarbituric reactive substances ( TBARS ) , nitric oxide levels ( NO ) , total radical antioxidant parameter ( TRAP ) , superoxide dismutase , and catalase activities and GSH levels . Immune evaluation was determined by TNF-α , IL-1β , IL-12 , and IL-10 levels and leukocytes oxidative burst evaluation by chemiluminescence . Tissue damage analysis included heart ( total CK and CKMB ) , liver ( AST , ALT , GGT ) , and renal ( creatinine , urea , and uric acid ) plasmatic markers . C-reactive protein ( CRP ) and iron metabolism were also evaluated . Analysis of the results verified different oxidative stress statuses occur at distinct cancer stages . ED was characterized by reduction in catalase , 8-isoprostanes , and GSH levels , with enhanced lipid peroxidation and TBARS levels . AD exhibited more pronounced oxidative status , with reduction in catalase activity and TRAP , intense lipid peroxidation and high levels of NO , TBARs , and carbonyl content . ED patients presented a Th2 immune pattern , while AD exhibited Th1 status . CRP levels and ferritin were increased in both stages of disease . Leukocytes burst impairment was observed in both the groups . Plasma iron levels were significantly elevated in AD . The data obtained indicated that oxidative stress enhancement and immune response impairment may be necessary to ensure cancer progression to advanced stages and may result from both host and tumor inflammatory mediators . OUTPUT: tumor promoting inflammation INPUT: We examined the influence of the level of dietary protein or vitamin E ( VE ) on oxidative damage to DNA , lipids , and protein in the liver after total body irradiation ( TBI ) with X-rays at 1 or 4 Gy . Levels of 8-hydroxydeoxyguanosine , thiobarbituric acid-reactive substances , and protein carbonyls in the liver did not differ among the groups that did not receive TBI . However , oxidative damage to lipids and protein was increased by TBI only in the 1% protein group . DNA damage , lipid peroxidation , or protein oxidation in the liver was increased by TBI in a dose-dependent manner , and the damage was consistently higher in the 1% than in the 20% protein group . In the 1% protein group , a greater decrease in relative spleen weight by TBI was also observed . Concentrations of antioxidants ( vitamins C and E and glutathione ) in the liver were lower and the concentration of nonheme iron in the liver was higher in the 1% than in the 20% protein group . Mice fed a 1% protein diet became susceptible to TBI-induced oxidative damage , and decreases in antioxidant levels and an increase in iron level were involved in the mechanism of this susceptibility . These results suggest that dietary VE and protein can prevent oxidative damage to DNA , lipid , and protein in mice subjected to TBI . Consumption of a VE-free diet significantly increased 8-hydroxydeoxyguanosine levels in DNA from mice fed the 1% protein diet with TBI , but such changes were not detected in DNA from mice fed the 20% protein diet . OUTPUT:

tumor promoting inflammation;genomic instability and mutation

HoC_dynamic_5_shot58

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Hepatocyte growth factor ( HGF ) , a mesenchymal-derived factor which regulates growth , motility , and morphogenesis of epithelial and endothelial cells , functions as a hepatotrophic and renotrophic factor for regeneration of the liver and kidney . We have now obtained evidence that transforming growth factor-beta 1 ( TGF-beta 1 ) and glucocorticoids are negative regulators for HGF gene expression . When TGF-beta 1 or dexamethasone was added to cultures of MRC-5 human embryonic lung fibroblasts and HL-60 human promyelocytic leukemic cells , the amount of HGF secreted into the culture medium was inhibited to 30-40% of that of control cultures by 10 ng/ml TGF-beta 1 and to 40-50% by 10(-6) M dexamethasone . The inhibitory effect of TGF-beta 1 and dexamethasone on HGF synthesis in MRC-5 cells was additive , thereby suggesting that TGF-beta 1 and dexamethasone exert effects through distinct mechanisms . Hydrocortisone also inhibited HGF synthesis with the same potency as dexamethasone ; however , testosterone , estriol , and beta-estradiol had no effect . The rate of HGF synthesis in MRC-5 cells , as measured by pulse labeling with [ 35S]methionine and subsequent immunoprecipitation , was suppressed to 30-40% of the control with 10 ng/ml TGF-beta 1 , and to 30-45% by 10(-6) M dexamethasone . HGF mRNA levels in MRC-5 cells and HL-60 cells were dose-dependently suppressed by TGF-beta 1 and dexamethasone ; 10 ng/ml TGF-beta 1 suppressed HGF mRNA levels to 32% and 35% of control culture , respectively , in MRC-5 cells and HL-60 cells , and 10(-6) M dexamethasone suppressed to 43% and 38% , respectively . Thus , TGF-beta 1 and glucocorticoids seem to inhibit HGF synthesis by suppressing the expression of the HGF gene . We propose that a negative regulation of HGF gene expression by TGF-beta 1 or glucocorticoids may be involved in physiological or pathological processes during tissue regeneration . OUTPUT: sustaining proliferative signaling INPUT: Melittin ( 1 ) is a major polypeptide in honey bee venom that has been used traditionally against chronic inflammation and cancer . However , its molecular mechanism has not been determined . In this study , the antitumor effect of 1 was compared with that of NS398 , a cyclooxygenase-2 ( COX-2 ) inhibitor , in vivo and in vitro . Subcutaneous injection of 1 at 0.5 and 5 mg/kg suppressed significantly vascular endothelial growth factor ( VEGF)-A-transfected highly metastatic Lewis lung cancer ( VEGF-A-hm LLC ) tumor growth by 25% and 57% , respectively . Also , 1 inhibited significantly the number of vessels around VEGF-A-hm LLC cells . The results were superior to those obtained in the mice treated with NS398 . Compound 1 dose-dependently inhibited proliferation and tube formation in human umbilical vein endothelial cells ( VEGF-A-HUVECs ) , without affecting cell viability in native HUVECs . In addition , 1 decreased the expression of VEGF receptor-2 ( VEGFR-2 ) , COX-2 , and prostaglandin E(2) ( PGE(2) ) in VEGF-A-transfected HUVECs . These effects were accompanied by a reduction of the phosphorylation of extracellular signal-regulated kinase 1/2 and c-jun N-terminal kinase , whereas it increased the phosphorylation of p38 mitogen-activated protein kinase ( MAPK ) . SB203580 abolished the downregulation of COX-2 and VEGFR-2 and the inhibition of cell proliferation by 1 . The antitumor activity of 1 may be associated with antiangiogenic actions via inhibiting VEGFR-2 and inflammatory mediators involved in the MAPK signaling pathway . OUTPUT: inducing angiogenesis;sustaining proliferative signaling INPUT: This study aimed to analyze the role of endothelial progenitor cell ( EPC)-derived angiogenic factors and chemokines in the multistep process driving angiogenesis with a focus on the recently discovered macrophage migration inhibitory factor ( MIF)/chemokine receptor axis . Primary murine and murine embryonic EPCs ( eEPCs ) were analyzed for the expression of angiogenic/chemokines and components of the MIF/CXC chemokine receptor axis , focusing on the influence of hypoxic versus normoxic stimulation . Hypoxia induced an upregulation of CXCR2 and CXCR4 but not CD74 on EPCs and triggered the secretion of CXCL12 , CXCL1 , MIF , and vascular endothelial growth factor ( VEGF ) . These factors stimulated the transmigration activity and adhesive capacity of EPCs , with MIF and VEGF exhibiting the strongest effects under hypoxia . MIF- , VEGF- , CXCL12- , and CXCL1-stimulated EPCs enhanced tube formation , with MIF and VEGF exhibiting again the strongest effect following hypoxia . Tube formation following in vivo implantation utilizing angiogenic factor-loaded Matrigel plugs was only promoted by VEGF . Coloading of plugs with eEPCs led to enhanced tube formation only by CXCL12 , whereas MIF was the only factor which induced differentiation towards an endothelial and smooth muscle cell ( SMC ) phenotype , indicating an angiogenic and differentiation capacity in vivo . Surprisingly , CXCL12 , a chemoattractant for smooth muscle progenitor cells , inhibited SMC differentiation . We have identified a role for EPC-derived proangiogenic MIF , VEGF and MIF receptors in EPC recruitment following hypoxia , EPC differentiation and subsequent tube and vessel formation , whereas CXCL12 , a mediator of early EPC recruitment , does not contribute to the remodeling process . By discerning the contributions of key angiogenic chemokines and EPCs , these findings offer valuable mechanistic insight into mouse models of angiogenesis and help to define the intricate interplay between EPC-derived angiogenic cargo factors , EPCs , and the angiogenic target tissue . OUTPUT: inducing angiogenesis INPUT: Heparin-binding epidermal growth factor-like growth factor ( HB-EGF ) is a member of the epidermal growth factor family and has a variety of physiological and pathological functions . Modulation of HB-EGF activity might have a therapeutic potential in the oncology area . We explored the therapeutic possibilities by characterizing the in vitro biological activity of anti-HB-EGF monoclonal antibody Y-142 . EGF receptor ( EGFR ) ligand and species specificities of Y-142 were tested . Neutralizing activities of Y-142 against HB-EGF were evaluated in EGFR and ERBB4 signaling . Biological activities of Y-142 were assessed in cancer cell proliferation and angiogenesis assays and compared with the anti-EGFR antibody cetuximab , the HB-EGF inhibitor CRM197 , and the anti-vascular endothelial growth factor ( VEGF ) antibody bevacizumab . The binding epitope was determined with alanine scanning . Y-142 recognized HB-EGF as well as the EGFR ligand amphiregulin , and bound specifically to human HB-EGF , but not to rodent HB-EGF . In addition , Y-142 neutralized HB-EGF-induced phosphorylation of EGFR and ERBB4 , and blocked their downstream ERK1/2 and AKT signaling . We also found that Y-142 inhibited HB-EGF-induced cancer cell proliferation , endothelial cell proliferation , tube formation , and VEGF production more effectively than cetuximab and CRM197 and that Y-142 was superior to bevacizumab in the inhibition of HB-EGF-induced tube formation . Six amino acids in the EGF-like domain were identified as the Y-142 binding epitope . Among the six amino acids , the combination of F115 and Y123 determined the amphiregulin cross-reactivity and that F115 accounted for the species selectivity . Furthermore , it was suggested that the potent neutralizing activity of Y-142 was derived from its recognition of R142 and Y123 and its high affinity to HB-EGF . Y-142 has a potent HB-EGF neutralizing activity that modulates multiple biological activities of HB-EGF including cancer cell proliferation and angiogenic activities . Y-142 may have a potential to be developed into a therapeutic agent for the treatment of HB-EGF-dependent cancers . OUTPUT: inducing angiogenesis;sustaining proliferative signaling INPUT: Histone deacetylase inhibitors ( HDACi ) are promising epigenetic cancer chemotherapeutics rapidly approaching clinical use . In this study , we tested using in vitro and in vivo models the differential biological effects of a novel HDAC inhibitor [ belinostat ( PXD101) ] , in a wide panel of androgen-sensitive and androgen-independent tumor cells . Belinostat significantly increased acetylation of histones H3 and H4 . Belinostat potently inhibited the growth of prostate cancer cell lines ( IC50 range from 0.5 to 2.5 �M ) with cytotoxic activity preferentially against tumor cells . This agent induced G2/M arrest and increased significantly the percentage of apoptosis mainly in androgen-sensitive tumor cells confirming its growth-inhibitory effects . The cell death mechanisms were studied in three different prostate cancer cell lines with different androgen dependence and expression of androgen receptor ; LAPC-4 and 22rv1 ( androgen-dependent and expressing androgen receptor ) and PC3 ( androgen-independent not expressing androgen receptor ) . Belinostat induced the expression of p21 and p27 , acetylation of p53 and G2/M arrest associated with Bcl2 and Bcl-Xl downmodulation and significant reduction of survivin , IAPs and Akt/pAkt and increased caspase-8 and -9 expression/activity . Belinostat effectiveness was dependent on the androgen receptor ( AR ) , since the stable transfection of AR greatly increased the efficacy of this HDAC inhibitor . These observations were correlated using in vivo models . We demonstrated that belinostat preferentially resulted in antitumor effect in androgen-dependent tumor cells expressing AR . Our findings provide evidence that belinostat may be a promising anticancer drug for prostate cancer expressing AR , supporting its clinical role in prostate cancer . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Expression of angiogenic factors is upregulated in hyperplastic mucosa adjacent to colon cancer , and this upregulation is closely associated with cancer growth and metastasis . We investigated the role of histone acetylation in vascular endothelial growth factor ( VEGF ) expression in hyperplastic mucosa adjacent to orthotopic colon cancer in mice . In the hyperplastic mucosa adjacent to KM12SM tumor in the cecum of athymic mice , VEGF upregulation was associated with hypoxia-inducible factor ( HIF)-1alpha induction . The hyperplastic mucosa also showed hypoacetylation of histone H4 and reduction of both p53 and von Hippel-Lindau ( VHL ) proteins . To examine the effects of growth factors and cytokines on histone acetylation and levels of p53 , VHL and HIF-1alpha , the rat intestinal epithelial cell line IEC6 was treated with epidermal growth factor ( EGF ) and interleukin ( IL)-15 for 35 days . Acetylated histone H4 , p53 protein and ubiquitinated protein levels were reduced , whereas HIF-1alpha production was upregulated in EGF- and IL-15-treated IEC6 cells . These findings suggest that EGF- or IL-15-induced histone H4 hypoacetylation is associated with repression of p53 and VHL genes in intestinal epithelial cells . The subsequent suppression of protein ubiquitination leads to upregulation of VEGF production by HIF-1alpha retention . OUTPUT:

inducing angiogenesis;sustaining proliferative signaling

HoC_dynamic_5_shot59

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: γ-Tocotrienol and sesamin are phytochemicals that display potent anticancer activity . Since sesamin inhibits the metabolic degradation of tocotrienols , studies were conducted to determine if combined treatment with sesamin potentiates the antiproliferative effects of γ-tocotrienol on neoplastic mouse ( +SA ) and human ( MCF-7 and MDA-MB-231 ) mammary cancer cells . Results showed that treatment with γ-tocotrienol or sesamin alone induced a significant dose-responsive growth inhibition , whereas combination treatment with these agents synergistically inhibited the growth of +SA , MCF-7 and MDA-MB-231 mammary cancer cells , while similar treatment doses were found to have little or no effect on normal ( mouse CL-S1 and human MCF-10A ) mammary epithelial cell growth or viability . However , sesamin synergistic enhancement of γ-tocotrienol-induced anticancer effects was not found to be mediated from a reduction in γ-tocotrienol metabolism . Rather , combined treatment with subeffective doses of γ-tocotrienol and sesamin was found to induce G1 cell cycle arrest , and a corresponding decrease in cyclin D1 , CDK2 , CDK4 , CDK6 , phospho-Rb , and E2F1 levels , and increase in p27 and p16 levels . Additional studies showed that the antiproliferative effect of combination treatment did not initiate apoptosis or result in a decrease in mammary cancer cell viability . Taken together , these findings indicate that the synergistic antiproliferative action of combined γ-tocotrienol and sesamin treatment in mouse and human mammary cancer cells is cytostatic , not cytotoxic , and results from G1 cell cycle arrest . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: We have previously demonstrated the antiproliferative effect of two flavonoids-2,2'-dihydroxychalcone ( DHC ) , a novel synthetic flavonoid , and fisetin , a naturally occurring flavonol-in prostate cancer cells . In this study , we further examine the mechanisms of these compounds on survival and proliferation pathways . DHC and fisetin ( 1-50 microM ) caused a dose-dependent reduction in viability , a concomitant increase in apoptosis in PC3 cells at 72 h , and a decrease in clonogenic survival at 24 h treatment . DHC was considerably more potent than fisetin in these cytotoxicity assays . The mechanism of accelerated cellular senescence was not activated by either compound in PC3 or lymph node carcinoma of the prostate ( LNCaP ) cells . Gene expression alterations in PC3 and LNCaP cells treated with 15 muM DHC and 25 microM fisetin for 6 to 24 h were determined by oligonucleotide array . Amongst the most highly represented functional categories of genes altered by both compounds was the cell cycle category . In total , 100 cell cycle genes were altered by DHC and fisetin including 27 genes with key functions in G2/M phase that were downregulated by both compounds . Other functional categories altered included chromosome organization , apoptosis , and stress response . These results demonstrate the multiple mechanisms of antitumor activity of DHC and fisetin in prostate cancer cells in vitro . OUTPUT: enabling replicative immortality;evading growth suppressors;sustaining proliferative signaling INPUT: The cytotoxic activity of cyclosporin A ( CsA ) and the three non-immuno-suppressive CsA analogues B3-243 , WO-039 and B3-665 were studied in tumor cell lines representing both classical and atypical forms of multidrug resistance ( MDR ) : T-ALL GM3639 L100 cells selected for vincristine ( vcr ) resistance and displaying characteristics of classical MDR , including P-glycoprotein ( pgp ) expression and increased drug efflux which can be inhibited by pgp blockers ( e.g. verapamil ) , and U-1285/ADR , a small cell lung cancer ( SCLC ) cell line selected for doxorubicin resistance which lacks pgp , is insensitive to pgp-blockers and shows cross resistance to cis-platinum . At 1 micrograms/ml CsA was the most active agent in reversing Vcr resistance in L100 cells followed by B3-243 and WO-039 , with no effect of B3-665 . Parental LO cells were only marginally sensitized to Vcr by these agents . No reversing effect of any cyclosporin was observed in the U-1285/ADR or its parental cell line . Compared to LO cells , L100 cells showed a marked hypersensitivity to CsA &gt ; B3-243 &gt ; WO-039 with B3-665 being inactive . No collateral sensitivity was observed for cyclosporins in U-1285/ADR cells . Although of different magnitude , the pattern of cytotoxic activity for the different cyclosporins alone closely parallelled that of L100 cells for U-1285 , U1285/ADR and LO cells . The results indicate that not only the collateral sensitivity in classical MDR but also the cytotoxic actions of cyclosporins per se on tumor cells alone are independent of immunosuppressive activity . The results also suggest a structure-activity relationship for cyclosporin-induced cytotoxicity similar to , but independent of , MDR reversing activity . OUTPUT: avoiding immune destruction INPUT: Evaluation of immune dysfunction during the tumor-bearing state is a critical issue in combating cancer . In this study , we initially found that IL-6 , one of the cachectic factors , suppressed CD4(+) T cell-mediated immunity through downregulation of MHC class II by enhanced arginase activity of dendritic cells ( DC ) in tumor-bearing mice . We demonstrated that administration of Ab against IL-6R ( anti-IL-6R mAb ) greatly enhanced T cell responses and inhibited the growth of tumor in vivo . We also found that IL-6 upregulated the expression of arginase-1 and arginase activity of DC in vitro . Tumor-infiltrating CD11c(+) DC exhibited upregulated mRNA expression of arginase-1 but reduced expression of MHC class II in parallel with the increase in serum IL-6 levels at the late stage in tumor-bearing hosts . However , the administration of anti-IL-6R mAb into tumor-bearing mice inhibited both the downmodulation of MHC class II and the upregulation of arginase-1 mRNA levels in DC . Furthermore , we noted that N(ω)-hydroxy-L-arginine or L-arginine , an arginase-1 inhibitor , blocked the reduction in MHC class II levels on CD11c(+) DC during the tumor-bearing state . Finally , we demonstrated that the administration of N(ω)-hydroxy-L-arginine at the peritumor site significantly enhanced CD4(+) T cell responses and inhibited tumor growth . Thus , IL-6-mediated arginase activation and the subsequent reduction in MHC class II expression on DC appeared to be critical mechanisms for inducing dysfunction of the immune system in the tumor-bearing state . Blockade of the IL-6-arginase cascade is a promising tool to overcome the dysfunction of antitumor immunity in tumor-bearing hosts . OUTPUT: avoiding immune destruction INPUT: The immune system has an important role in tumor appearance and spreading . One of the most efficient subpopulations of cytotoxic cells in the destruction of tumors are NK cells . NK cells are activated and increase their cytotoxic potential and modulate their cytokine production after treatment with IFNgamma , IL-12 , TNFalpha and IL-2 . The investigation of the activity of NK cells was performed on peripheral blood lymphocytes ( PBL ) of 16 healthy controls and of 40 patients with metastatic breast carcinoma . Modulation of NK cells was performed with IL-2 , IL-7 , IL-12 , TNFalpha , monoclonal antibodies ( mAb ) for TNFalpha and TNFalpha receptors type I and II , as well as with sera of healthy controls and patients with breast cancer in different clinical stages . Modulating effect of the applied factors after in vitro treatment of PBL was evaluated by the cytotoxic assay using 51chromium . Our results indicate that IL-2 significantly increased the activity of NK cells of controls and breast cancer patients . The sera of patients with advanced breast cancer significantly reduced NK cell activity . IL-7 , IL-12 and mAb for TNFalpha do not significantly change the activity of NK cells . The presence of anti-TNFalpha mAb did not change the inhibitory effect of the sera of breast cancer patients with advanced disease on the activity of NK cells of controls and patients with breast cancer . Blocking of TNFalpha Rcs with mAbs decrease the reactivity of NK cells for IL-2 . The treatment of breast cancer patients with advanced clinical stage of breast cancer with IL-2 , as an additional therapy , could be advantageous , as NK cells after this treatment increase their cytotoxic activity against tumor cells and can improve therapeutical results . OUTPUT: avoiding immune destruction INPUT: The effects of an anti-CD3 mAb on induction of non-MHC restricted cytolysis was investigated . Peripheral blood mononuclear cells ( PBMC ) from normal donors ( 29 ) and cancer patients ( 18 ) were cultured in 100 U/ml of interleukin-2 ( rIL-2 ) with and without anti-CD3 mAb ( OKT3 , 10 ng/ml ) for the first 48 hours of incubation . Thereafter , both PBMC cultures were maintained on rIL-2 up to 20 days . PBMC proliferation was enhanced 17-fold in number by day 20 when anti-CD3 mAb and rIL-2 was present during the first 48 hours but only 3-fold by day 20 when rIL-2 alone was present . Concomitantly anti-CD3 mAb but not Lym-1 , an isotype matched control , inhibited the induction of lytic activity against both NK sensitive ( K562 ) and NK resistant ( Raji ) target cell lines . Thus the inhibitory effect is dependent on anti-CD3 mAb stimulating the CD3/TCR T-cell receptor complex . While lytic activity was dependent on the concentration of rIL-2 , inhibition of the induction phase of non-MHC restricted lytic activity was independent of the concentration of rIL-2 . Flow cytometry analysis indicated that treatment with the anti-CD3 mAb increased the percentage of CD3 positive cells , CD4 positive cells and especially CD25 positive cells , but decreased th percentage of CD56 positive cells . Supernatants from anti-CD3 mAb stimulated cultures also inhibited the induction of non-MHC restricted lytic activity . Lymphokine analysis showed that supernatants of anti-CD3 mAb stimulated cultures had higher levels of TNF-alpha and IFN-gamma . However , TNF-alpha and IFN-gamma alone or in combination could not mediate the inhibitory effect . The inhibitory factor(s) was partially purified by sequential chromatography on matrices of controlled pore glass and Sepharose CL-6B . The molecular weight of the inhibitory factor(s) was less than 67K . These studies have identified a novel regulatory pathway controlling non-MHC restricted cytolysis . Perturbation of the T-cell CD3/TCR complex with the anti-CD3 mAb results in the secretion of a soluble mediator that down-regulates the induction of rIL-2 dependent non-MHC restricted cytolysis . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot60

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Identification of the proteins that are associated with estrogen receptor ( ER ) status is a first step towards better understanding of the hormone-dependent nature of breast carcinogenesis . Although a number of gene expression analyses have been conducted , protein complement has not been systematically investigated to date . Because proteins are primary targets of therapeutic drugs , in this study , we have attempted to identify proteomic signatures that demarcate ER-positive and -negative breast cancers . Using highly enriched breast tumor cells , replicate analyses from 3 ERα+ and 3 ERα- human breast tumors resulted in the identification of 2,995 unique proteins with ≥2 peptides . Among these , a number of receptor tyrosine kinases and intracellular kinases that are abundantly expressed in ERα+ and ERα- breast cancer tissues were identified . Further , label-free quantitative proteome analysis revealed that 236 proteins were differentially expressed in ERα+ and ERα- breast tumors . Among these , 141 proteins were selectively up-regulated in ERα+ , and 95 proteins were selectively up-regulated in ERα- breast tumors . Comparison of differentially expressed proteins with a breast cancer database revealed 98 among these have been previously reported to be involved in breast cancer . By Gene Ontology molecular function , dehydrogenase , reductase , cytoskeletal proteins , extracellular matrix , hydrolase , and lyase categories were significantly enriched in ERα+ , whereas selected calcium-binding protein , membrane traffic protein , and cytoskeletal protein were enriched in ERα- breast tumors . Biological process and pathway analysis revealed that up-regulated proteins of ERα+ were overrepresented by proteins involved in amino acid metabolism , proteasome , and fatty acid metabolism , while up-regulated proteins of ERα- were overrepresented by proteins involved in glycolysis pathway . The presence and relative abundance of 4 selected differentially abundant proteins ( liprin-α1 , fascin , DAP5 , and β-arrestin-1 ) were quantified and validated by immunohistochemistry . In conclusion , unlike in vitro cell culture models , the in vivo signaling proteins and pathways that we have identified directly from human breast cancer tissues may serve as relevant therapeutic targets for the pharmacological intervention of breast cancer . OUTPUT: cellular energetics INPUT: The contribution that mitochondrial bioenergetics could have in cancer development is debated . Here , we have generated HCT116-derived colocarcinoma cell lines expressing different levels of the beta catalytic subunit of the mitochondrial H+-adenosine triphosphate synthase to assess the contribution of mitochondrial bioenergetics in colon cancer progression . The generated cells exhibit large ultrastructural , transcriptomic , proteomic and functional differences in their mitochondria and in their in vivo tumor forming capacity . We show that the activity of oxidative phosphorylation defines the rate of glucose utilization by aerobic glycolysis . The aggressive cellular phenotype , which is highly glycolytic , is bound to the deregulated expression of genes involved in metabolic processes , the regulation of the cell cycle , apoptosis , angiogenesis and cell adhesion . Remarkably , the molecular and ultrastructural analysis of the tumors derived from the three HCT116 cell lines under study highlight that tumor promotion inevitably requires the selection of cancer cells with a repressed biogenesis and functional activity of mitochondria , i.e. the highly glycolytic phenotype is selected for tumor development . The tumor forming potential of the cells is a non-genetically acquired condition that provides the cancer cell with a cell-death resistant phenotype . An abrogated mitochondrial respiration contributes to a diminished potential for reactive oxygen species signaling in response to 5-fluorouracil treatment . Treatment of cancer cells with dichloroacetate partially restores the functional differentiation of mitochondria and promotes tumor regression , emphasizing the reversible nature of the metabolic trait of cancer . OUTPUT: cellular energetics INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: BACKGROUND Lung cancer often develops in association with chronic pulmonary inflammatory diseases with an influx of neutrophils . More detailed information on inflammatory pathways and the role of neutrophils herein is a prerequisite for understanding the mechanism of inflammation associated cancer . METHODS In the present study , we used microarrays in order to obtain a global view of the transcriptional responses of the lung to LPS in mice , which mimics an acute lung inflammation . To investigate the influence of neutrophils in this process , we depleted mice from circulating neutrophils by treatment with anti-PMN antibodies prior to LPS exposure . RESULTS A total of 514 genes was greater than 1.5-fold differentially expressed in the LPS induced lung inflammation model. 394 of the 514 were up regulated genes mostly involved in cell cycle and immune/inflammation related processes , such as cytokine/chemokine activity and signalling . Down regulated genes represented nonimmune processes , such as development , metabolism and transport . Notably , the number of genes and pathways that were differentially expressed , was reduced when animals were depleted from circulating neutrophils , confirming the central role of neutrophils in the inflammatory response . Furthermore , there was a significant correlation between the differentially expressed gene list and the promutagenic DNA lesion M1dG , suggesting that it is the extent of the immune response which drives genetic instability in the inflamed lung . Several genes that were specifically regulated by the presence of activated neutrophils could be identified and these were mostly involved in interferon signalling , oxidative stress response and cell cycle progression . The latter possibly refers to a higher rate of cell turnover in the inflamed lung with neutrophils , suggesting that the neutrophil influx is associated with a higher risk for the accumulation and fixation of mutations . CONCLUSION Gene expression profiling identified specific genes and pathways that are related to neutrophilic inflammation and could be associated to cancer development and indicate an active role of neutrophils in mediating the LPS induced inflammatory response in the mouse lung . OUTPUT: tumor promoting inflammation;genomic instability and mutation INPUT: Progression to malignancy requires that cells overcome senescence and switch to an immortal phenotype . Thus , exploring the genetic and epigenetic changes that occur during senescence/immortalization may help elucidate crucial events that lead to cell transformation . In the present study , we have globally profiled DNA methylation in relation to gene expression in primary , senescent and immortalized mouse embryonic fibroblasts . Using a high-resolution genome-wide mapping technique , followed by extensive locus-specific validation assays , we have identified 24 CpG islands that display significantly higher levels of CpG methylation in immortalized cell lines as compared to primary murine fibroblasts . Several of these hypermethylated CpG islands are associated with genes involved in the MEK-ERK pathway , one of the most frequently disrupted pathways in cancer . Approximately half of the hypermethylated targets are developmental regulators , and bind to the repressive Polycomb group ( PcG ) proteins , often in the context of bivalent chromatin in mouse embryonic stem cells . Because PcG-associated aberrant DNA methylation is a hallmark of several human malignancies , our methylation data suggest that epigenetic reprogramming of pluripotency genes may initiate cell immortalization . Consistent with methylome alterations , global gene expression analysis reveals that the vast majority of genes dysregulated during cell immortalization belongs to gene families that converge into the MEK-ERK pathway . Additionally , several dysregulated members of the MAP kinase network show concomitant hypermethylation of CpG islands . Unlocking alternative epigenetic routes for cell immortalization will be paramount for understanding crucial events leading to cell transformation . Unlike genetic alterations , epigenetic changes are reversible events , and as such , can be amenable to pharmacological interventions , which makes them appealing targets for cancer therapy when genetic approaches prove inadequate . OUTPUT: enabling replicative immortality INPUT: Induction of the expression of the Mr 67,000 high-affinity laminin receptor gene has been postulated as playing a role in the progression of human tumors to invasive cancers . We tested this hypothesis by examining histopathological sections of a large number of epithelial lesions of the genital tract associated with human papillomaviruses . In situ hybridization was performed with a riboprobe generated from a laminin receptor complementary DNA . Laminin receptor mRNA was expressed primarily in the less differentiated cells in normal squamous tissues and in a spectrum of squamous neoplasms . There was no net induction of mRNA per cell in intraepithelial or invasive squamous neoplasms relative to normal tissue . In contrast , laminin receptor mRNA was not expressed at a detectable level in normal glands of the uterine cervix but was dramatically induced in morphologically abnormal , human papillomavirus-positive glands , irrespective of the genotype of human papillomaviruses present . The induction occurred before any evidence of invasion , and there was no further increase during the transition from adenocarcinoma in situ to invasive carcinoma . We conclude that induction of high-affinity laminin receptor gene expression is associated with the development of malignancies of cervical glandular epithelia , but the increased expression appears to correlate with the proliferative rather than the invasive properties of these cells . OUTPUT:

sustaining proliferative signaling;activating invasion and metastasis

HoC_dynamic_5_shot61

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Aims : RAS-induced tumorigenesis has been suggested to follow a three-stage model consisting of an initial RAS activation , senescence induction , and evasion of p53-dependent senescence checkpoints . While reactive oxygen species ( ROS ) act as second messengers in RAS-induced senescence , they are also involved in oncogenic transformation by inducing proliferation and promoting mutations . In the current work , we investigated the role of extracellular superoxide dismutase ( SOD3 ) in RAS-induced senescence and immortalization in vitro and in vivo . We used a mouse embryonic fibroblast ( MEF ) primary cell model together with immortalized and transformed human cell lines derived from papillary and anaplastic thyroid cancer . Results : Based on our data , sod3 RNA interference in H-RasV12-transduced cells markedly inhibited cell growth , while sod3 over-expression in MEFs initially caused a proliferative burst followed by the activation of DNA damage checkpoints , induction of p53-p21 signal transduction , and senescence . Subsequently , sod3-transduced MEF cells developed co-operative p21-p16 down-regulation and acquired transformed cell characteristics such as increased telomerase activity , loss of contact inhibition , growth in low-nutrient conditions , and in vivo tumorigenesis . Interestingly , as reported previously with RAS , we showed a dose-dependent response to SOD3 in vitro and in vivo involving transcriptional and non-transcriptional regulatory mechanisms . Innovation : SOD3 may mediate H-RasV12-induced initiation of primary cell immortalization . Conclusions : Our results indicate that SOD3 influences growth signaling in primary and cancer cells downstream of the ras oncogene and could serve as a therapy target at an early tumorigenesis phase . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;tumor promoting inflammation;sustaining proliferative signaling INPUT: Epigenetic changes in pediatric neuroblastoma may contribute to the aggressive pathophysiology of this disease , but little is known about the basis for such changes . In this study , we examined a role for the DNA methyltransferase DNMT3B , in particular , the truncated isoform DNMT3B7 , which is generated frequently in cancer . To investigate if aberrant DNMT3B transcripts alter DNA methylation , gene expression , and phenotypic character in neuroblastoma , we measured DNMT3B expression in primary tumors . Higher levels of DNMT3B7 were detected in differentiated ganglioneuroblastomas compared to undifferentiated neuroblastomas , suggesting that expression of DNMT3B7 may induce a less aggressive clinical phenotype . To test this hypothesis , we investigated the effects of enforced DNMT3B7 expression in neuroblastoma cells , finding a significant inhibition of cell proliferation in vitro and angiogenesis and tumor growth in vivo . DNMT3B7-positive cells had higher levels of total genomic methylation and a dramatic decrease in expression of the FOS and JUN family members that comprise AP1 transcription factors . Consistent with an established antagonistic relationship between AP1 expression and retinoic acid receptor activity , increased differentiation was seen in the DNMT3B7-expressing neuroblastoma cells following treatment with all-trans retinoic acid ( ATRA ) compared to controls . Our results indicate that DNMT3B7 modifies the epigenome in neuroblastoma cells to induce changes in gene expression , inhibit tumor growth , and increase sensitivity to ATRA . OUTPUT: inducing angiogenesis;sustaining proliferative signaling INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: SET and MYND domain-containing protein 3 ( SMYD3 ) is a histone methyltransferase that plays an important role in transcriptional regulation in human carcinogenesis . It can specifically methylate histone H3 at lysine 4 and activate the transcription of a set of downstream genes , including several oncogenes ( e.g. , N-myc , CrkL , Wnt10b , RIZ and hTERT ) and genes involved in the control of cell cycle ( e.g. , CyclinG1 and CDK2 ) and signal transduction ( e.g. , STAT1 , MAP3K11 and PIK3CB ) . To determine the effects of SMYD3 over-expression on cell proliferation , we transfected SMYD3 into MDA-MB-231 cells and found that these cells showed several transformed phenotypes as demonstrated by colony growth in soft agar . Besides , we show here that down-regulation of SMYD3 could induce G1-phase cell cycle arrest , indicating the potent induction of apoptosis by SMYD3 knockdown . These results suggest the regulatory mechanisms of SMYD3 on the acceleration of cell cycle and facilitate the development of strategies that may inhibit the progression of cell cycle in breast cancer cells . OUTPUT: evading growth suppressors;resisting cell death INPUT: To elucidate the function of MAS-related GPCR , member D ( MRGD ) in cancers , we investigated the in vitro and in vivo oncogenic function of MRGD using murine fibroblast cell line NIH3T3 in which MRGD is stably expressed . The expression pattern of MRGD in clinical samples was also analyzed . We found that overexpression of MRGD in NIH3T3 induced focus formation and multi-cellular spheroid formation , and promoted tumors in nude mice . In other words , overexpression of MRGD in NIH3T3 induced the loss of contact inhibition , anchorage-independent growth and in vivo tumorigenesis . Furthermore , it was found that the ligand of MRGD , beta-alanine , enhanced spheroid formation in MRGD-expressing NIH3T3 cells . From investigation of clinical cancer tissues , we found high expression of MRGD in several lung cancers by immunohistochemistry as well as real time PCR . Based on these results , MRGD could be involved in tumorigenesis and could also be a novel anticancer drug target . OUTPUT: evading growth suppressors INPUT: Mas oncogene has been shown to have focus-inducing ability in NIH 3T3 cells which are tumorigenic in vivo in nude mice . Its stable expression in a variety of cell lines conferred some angiotensin responsiveness . To understand why mas-transfected cells exhibit a transformed phenotype and if angiotensin responsiveness plays any role in this process , we studied the growth characteristics of mas-transfected 3T3 cells and demonstrated that they lose contact inhibition , exhibit foci formation , and increased DNA synthesis even in absence of serum . Our results suggest that the transformed phenotype is due to the production of a mas receptor ligand distinct from angiotensin . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot62

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Agricultural products and by products provide the primary materials for a variety of technological applications in diverse industrial sectors . Agro-industrial wastes , such as cotton and curaua fibers , are used to prepare nanofibers for use in thermoplastic films , where they are combined with polymeric matrices , and in biomedical applications such as tissue engineering , amongst other applications . The development of products containing nanofibers offers a promising alternative for the use of agricultural products , adding value to the chains of production . However , the emergence of new nanotechnological products demands that their risks to human health and the environment be evaluated . This has resulted in the creation of the new area of nanotoxicology , which addresses the toxicological aspects of these materials . PURPOSE AND METHODS Contributing to these developments , the present work involved a genotoxicological study of different nanofibers , employing chromosomal aberration and comet assays , as well as cytogenetic and molecular analyses , to obtain preliminary information concerning nanofiber safety . The methodology consisted of exposure of Allium cepa roots , and animal cell cultures ( lymphocytes and fibroblasts ) , to different types of nanofibers . Negative controls , without nanofibers present in the medium , were used for comparison . RESULTS The nanofibers induced different responses according to the cell type used . In plant cells , the most genotoxic nanofibers were those derived from green , white , and brown cotton , and curaua , while genotoxicity in animal cells was observed using nanofibers from brown cotton and curaua . An important finding was that ruby cotton nanofibers did not cause any significant DNA breaks in the cell types employed . CONCLUSION This work demonstrates the feasibility of determining the genotoxic potential of nanofibers derived from plant cellulose to obtain information vital both for the future usage of these materials in agribusiness and for an understanding of their environmental impacts . OUTPUT: genomic instability and mutation INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Immunosuppression has been related to the incidence of tumor apparition , including endocrine tumors . The intrasplenic ovarian tumor ( luteoma ) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and , from the immunological perspective , is located in a critical organ involved in immune response . To establish if immunosuppression could alter the development of this experimental tumor , the effects of cyclosporin A ( CsA ) and dexamethasone ( Dex ) were evaluated . After surgery , tumor-bearing and sham animals were kept without treatment for 4 weeks ; thereafter , they were distributed into CsA ( 25 mg/kg ) , Dex ( 0.1 mg/kg ) , or vehicle ( 75:25 castor oil:ethanol ) groups and were injected on alternate days for 50 days . Body weight was evaluated weekly . Animals were sacrificed after a jugular vein blood sample was obtained . Thymi were weighed . Tumors were measured and placed in formaline for histological studies . Serum luteinizing hormone ( LH ) , follicle-stimulating hormone ( FSH ) , prolactin ( PRL ) , and estradiol were measured by radioimmunoassay . Hematological parameters were determined . CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals ( P &lt ; 0.01 ) . Dex significantly impaired weight increase in both groups of animals . CsA induced a significant weight loss in sham animals , not observed in tumor-bearing animals . Dex induced thymus weight loss in both groups , whereas CsA induced thymus weight loss only in sham animals . Only Dex induced a decrease in lymphocyte number in both groups . CsA induced an increase in monocyte number only in sham animals . Treatments did not alter LH , FSH , or estradiol , whereas PRL was increased by CsA only in sham rats . Neither Dex nor CsA induced any significant variations in tumor volume , nor did they alter tumor histology . In addition , no visible metastases or alterations in other organs were observed . We conclude that , though immunological parameters were altered by the treatments , immunosuppressor drugs did not condition tumor development . In addition , tumors secrete one or more factor/s that counteract CsA effect . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: BACKGROUND Engineered zinc-finger nucleases ( ZFN ) represented an innovative method for the genome manipulation in vertebrates . ZFN introduced targeted DNA double strand breaks ( DSB ) and initiated non-homologous end joining ( NHEJ ) after pronuclear or cytoplasmatic microinjection into zygotes . Resulting frame shift mutations led to functional gene ablations in zebra fish , mice , pigs and also in laboratory rats . Therefore , we targeted the rat Rag1 gene essential for the V(D)J recombination within the immunoglobulin production process and for the differentiation of mature B and T lymphocytes to generate an immunodeficient rat model in the LEW/Ztm strain . RESULTS After microinjection of Rag1 specific ZFN mRNAs in 623 zygotes of inbred LEW/Ztm rats 59 offspring were born from which one carried a 4 bp deletion . This frame shift mutation led to a premature stop codon and a subsequently truncated Rag1 protein confirmed by the loss of the full-length protein in Western Blot analysis . Truncation of the Rag1 protein was characterized by the complete depletion of mature B cells . The remaining T cell population contained mature CD4+/CD3+/TCRαβ+ as well as CD8+/CD3+/TCRαβ+ positive lymphocytes accompanied by a compensatory increase of natural killer cells in the peripheral blood . Reduction of T cell development in Rag1 mutant rats was associated with a hypoplastic thymus that lacked follicular structures . Histological evaluation also revealed the near-complete absence of lymphocytes in spleen and lymph nodes in the immunodeficient Rag1 mutant rat . CONCLUSION The Rag1 mutant rat will serve as an important model for transplantation studies . Furthermore , it may be used as a model for reconstitution experiments related to the immune system , particularly with respect to different populations of human lymphocytes , natural killer cells and autoimmune phenomena . OUTPUT: genomic instability and mutation;avoiding immune destruction INPUT: The mouse polyomavirus encodes a tumor-suppressor gene inactivator in its large T protein and a proto-oncogene activator in its middle T protein . We have used site-directed mutagenesis to selectively inactivate the former function without affecting the latter . Two mutant viruses were constructed to encode altered large T proteins that fail to bind the retinoblastoma tumor-suppressor gene product pRB , along with normal small and middle T proteins . The pRB-binding mutants proved to be defective in immortalization of primary rat embryo fibroblasts by a variety of tests . Yet they proved capable of transforming both primary and established fibroblasts in culture . Most importantly , the inability of these mutants to bind pRB had little effect on their ability to induce tumors in mice . We conclude that induction of multiple tumor types in this system does not depend on large T-pRB interactions but rather on middle T-dependent pathways . In addition , the ability of this virus to immortalize cells in culture is not essential to its ability to induce tumors in the animal . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot63

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Although tumor-associated macrophages ( TAMs ) are involved in tumor growth and metastasis , the mechanisms controlling their pro-tumoral activities remain largely unknown . The transcription factor c-MYC has been recently shown to regulate in vitro human macrophage polarization and be expressed in macrophages infiltrating human tumors . In this study , we exploited the predominant expression of LysM in myeloid cells to generate c-Myc(fl/fl) LysM(cre/+) mice , which lack c-Myc in macrophages , to investigate the role of macrophage c-MYC expression in cancer . Under steady-state conditions , immune system parameters in c-Myc(fl/fl) LysM(cre/+) mice appeared normal , including the abundance of different subsets of bone marrow hematopoietic stem cells , precursors and circulating cells , macrophage density , and immune organ structure . In a model of melanoma , however , TAMs lacking c-Myc displayed a delay in maturation and showed an attenuation of pro-tumoral functions ( e.g. , reduced expression of VEGF , MMP9 , and HIF1α ) that was associated with impaired tissue remodeling and angiogenesis and limited tumor growth in c-Myc(fl/fl) LysM(cre/+) mice . Macrophage c-Myc deletion also diminished fibrosarcoma growth . These data identify c-Myc as a positive regulator of the pro-tumoral program of TAMs and suggest c-Myc inactivation as an attractive target for anti-cancer therapy . OUTPUT: avoiding immune destruction INPUT: Previous reports showed that treatment with non-steroidal anti-inflammatory agents ( NSAIA ) can alter the growth profile of a variety of tumours . In this study , the effect of NSAIA treatment on the growth of the primary tumour and the appearance of spontaneous pulmonary metastases , was investigated . A mammary adenocarcinoma of non-detected immunogenicity , C7HI , was grafted subcutaneously in the lateral flank of Balb/c mice . Oral treatment with approximately 1 mg kg-1 day-1 piroxicam delayed both tumour growth and the growth of pulmonary metastases . Survival of mice bearing the primary tumour was significantly lengthened by anti-inflammatory treatment . Similarly , in separate experiments , after surgical removal of the primary tumour by day 34 after grafting , the group of mice treated orally with piroxicam also exhibited a higher survival rate than the control group . Upon surgical removal of the primary tumour 34 days after grafting , piroxicam treatment significantly decreased both the number and size of pulmonary metastases . The results of this study lends support to the hypothesis that inhibition or modulation of inflammation may delay tumour organisation and growth . It is suggested that piroxicam treatment may be an appropriate adjunct therapy to delay the appearance of pulmonary metastases and to increase life-expectancy in a host whose primary tumour has to be surgically removed . OUTPUT: activating invasion and metastasis;tumor promoting inflammation INPUT: Hormone-independent tumor growth and metastasis are associated with increased mortality in human prostate cancer . In this study , we evaluate a potential role for ligand-mediated activation of HER2 receptor tyrosine kinase in androgen-independent prostate cancers . HER2 , HER3 , and epidermal growth factor receptor were detected in the androgen-independent cell line 22Rv1 . Heregulin stimulation results in receptor phosphorylation and cell proliferation that is inhibited by increasing concentrations of anti-HER2 recombinant humanized monoclonal antibody ( rhuMAb ) 2C4 . Furthermore , inhibition of tumor growth was observed in xenografts derived from 22Rv1 cells when treated with rhuMAb 2C4 in a dose-dependent manner . These studies provide a framework , both in vitro and in vivo , to examine the molecular mechanisms of ligand-driven HER2 activation in androgen-independent tumorigenesis . OUTPUT: sustaining proliferative signaling INPUT: Hormone-dependent estrogen receptor positive ( ER+ ) breast cancers generally respond well to anti-estrogen therapy . Unfortunately , hormone-independent estrogen receptor negative ( ER- ) breast cancers are aggressive , respond poorly to current treatments and have a poor prognosis . New approaches and targets are needed for the prevention and treatment of ER- breast cancer . The NF-κB signaling pathway is strongly implicated in ER- tumor genesis , constituting a possible target for treatment . Hydrogen sulfide-releasing aspirin ( HS-ASA ) , a novel and safer derivative of aspirin , has shown promise as an anti-cancer agent . We examined the growth inhibitory effect of HS-ASA via alterations in cell proliferation , cell cycle phase transitions , and apoptosis , using MDA-MB-231 cells as a model of triple negative breast cancer . Tumor xenografts in mice , representing human ER- breast cancer , were evaluated for reduction in tumor size , followed by immunohistochemical analysis for proliferation , apoptosis and expression of NF-κB . HS-ASA suppressed the growth of MDA-MB-231 cells by induction of G(0)/G(1) arrest and apoptosis , down-regulation of NF-κB , reduction of thioredoxin reductase activity , and increased levels reactive oxygen species . Tumor xenografts in mice , were significantly reduced in volume and mass by HS-ASA treatment . The decrease in tumor mass was associated with inhibition of cell proliferation , induction of apoptosis and decrease in NF-κB levels in vivo . HS-ASA has anti-cancer potential against ER- breast cancer and merits further study . OUTPUT: resisting cell death;sustaining proliferative signaling;tumor promoting inflammation INPUT: BACKGROUND The most deadly form of cancer is not lung or colon , breast or prostate ; it is any cancer that has become metastatic . Mortality due to metastatic melanoma , one of the most aggressive and deadly cancers , has increased steadily over the last several decades . Unfortunately , the arsenal of chemotherapeutic agents available today is most often unsuccessful at extending and improving the life expectancy of afflicted individuals . We sought to identify an effective and nontoxic agent against metastatic melanoma . METHODOLOGY/PRINCIPAL FINDINGS We chose to study Cloudman S-91 mouse melanoma cells ( sub-clone M3 , CCL53.1 ) because these cells are highly aggressive and metastatic , representing one of the deadliest types of cancer . Melanoma cells also had an experimental advantage because their morphology , which is easily monitored , relates to the physiology of metastatic cells and normal melanocytes . We chose to test methyl sulfone as a chemotherapeutic agent for two reasons . Because of its chemical structure , we speculated a potential anti-cancer activity by targeting microtubules . Equally important , methyl sulfone has a well-established safety profile in humans . Surprisingly , we found that malignant melanoma cells exposed to methyl sulfone demonstrated the loss of phenotypes characteristic of malignant cells , and the reemergence of phenotypes characteristic of healthy melanocytes . Briefly , over time methyl sulfone induced contact inhibition , loss of ability to migrate through an extracellular matrix , loss of anchorage-independent growth , proper wound healing followed by contact inhibition , irreversible senescence followed by arborization with melanosomes in arbors as seen in normal melanocytes . CONCLUSIONS/SIGNIFICANCE Methyl sulfone may have clinical potential as a non-toxic agent effective against metastatic melanoma . Additionally , methyl sulfone has promise as a tool to explore molecular mechanisms of metastatic transformation as well as fundamental processes such as cell migration , contact inhibition , wound healing and cellular senescence . OUTPUT: enabling replicative immortality;evading growth suppressors;activating invasion and metastasis INPUT: Melanocyte stimulating hormone ( alpha-MSH , alpha-melanotropin),Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Ly-Pro-Va l-NH2 , regulates melanogenesis within epidermal melanocytes of many animals . An MSH analogue ( [ Nle4,D-Phe7]alpha-MSH ) that exhibits superpotency and prolonged biological activity has been synthesized , biologically characterized , and is presently in clinical trials to determine its possible clinical use in tanning of the skin . It also has potential for the diagnosis , localization , and chemotherapy of melanoma . The effects of this analogue on the growth , metastatic behavior , and invasive potential of a melanotic variant of Cloudman S-91 murine melanoma are reported here . In an intracutaneous murine model of melanoma cell tumor growth , the analogue did not increase primary tumor growth ( size ) after the period of administration of the peptide hormone analogue and did not affect spontaneous lung metastases . Survival times for the control and melanotropin-treated groups were similar , suggesting that overall tumor burden was not affected by treatment with the hormone analogue . Last , melanoma cell invasion through a human amniotic basement membrane in vitro was not enhanced compared to untreated cells . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot64

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: It is well known that cell-mediated immunity is suppressed in patients with neoplastic diseases . We have reported that soluble receptors for interleukin-2 ( sIL-2R ) and tumor necrosis factor ( sTNF-R1 ) are elevated in the serum of patients with advanced colorectal cancer . The presence of these soluble receptors and immunosuppressive cytokines , including interleukin-10 ( IL-10 ) , might be important in the mechanisms of immunosuppression. cis-Diaminedichloroplatinum ( cisplatin ) has been reported to immunomodulate , especially when used in low dose in combination with 5-Fluorouracil ( 5-FU ) . In this study , cisplatin and UFT , a form of uracil and tegafur which is a prodrug of 5-FU , were administered with immunomodulator Polysaccharide K ( PSK ) to ten patients with colorectal cancer , who showed distant metastasis in the liver or lung , and the serum levels of sIL-2R and sTNF-R1 and the production of gamma-interferon ( gamma-INF ) and interleukin-10 by peripheral blood mononuclear cells were measured . The serum concentrations of sIL-2R and the production of IL-10 were reduced ( p &lt ; 0.05 ) after 2 months of treatment . Thus , this combination appeared to have immunomodulative potential in patients with advanced colorectal cancer . OUTPUT: activating invasion and metastasis;avoiding immune destruction INPUT: In order to determine the in vivo immune response in glioblastoma , monoclonal and polyclonal antibodies specific for inflammatory leukocytes and immunoregulatory products were utilized to stain tissue from four surgical specimens . The more activated the inflammatory cells , the more activated the tumors appeared to be . In the tumor with the largest infiltration ( Case 3 ) , inflammatory cells were stained for interferon-gamma , interleukin-2 , interleukin-1 beta , lymphotoxin , tumor necrosis factor-alpha , and transforming growth factor-beta . The tumor cells also expressed interleukin-1 beta , interleukin-6 , transforming growth factor-beta , tumor necrosis factor-alpha , and prostaglandin E. In contrast , in the tumor with the least inflammatory response ( Case 1 ) , the tumor cells did not express any cytokines . Expression of cytokines by glioma cells was modest in the two cases with modest inflammatory responses . Cellular inflammation , primarily consisting of T cells and macrophages with few or no B cells or natural killer cells , was two- to 15-fold greater outside the tumor than within . In contrast to leukocytes outside the tumor , which were activated and expressing class II major histocompatibility antigens , leukocytes within the tumor parenchyma or at the tumor's edge were negative for these antigens . In the four specimens studied here , the tumor cells themselves were also negative for class II major histocompatibility antigens . These findings , although preliminary , suggest that inflammatory cells within gliomas are inactivated and that glioma cells may increase the expression of immunosuppressive cytokines in response to an increased lymphocyte infiltrate . This observation , if corroborated by more extensive studies , may help to explain the failure of immune treatments in glioblastoma multiforme . OUTPUT: avoiding immune destruction;tumor promoting inflammation INPUT: Immunosenescence , the progressive decline of adaptive immunity and chronic inflammation with ageing has been demonstrated to be the main factor responsible for infections , cancer and autoimmune conditions in the elderly . Senescence-accelerated mouse ( SAM ) was used to study the protective effects of Pu-erh tea in the elderly . The senile-prone sub-strain , SAM-P8 mice were administered individually with ripened or crude Pu-erh tea at 125 , 250 or 500mg/kg . The results showed that Pu-erh tea significantly increased the fractions of na�ve T lymphocytes , CD8(+)CD28(+) T lymphocytes and NK cells in the peripheral blood , but decreased the levels of IL-6 in aged mice . These data suggested that the Pu-erh tea reversed the immunosenescence by restoring the immune deficiency and decreasing pro-inflammatory cytokine . Thus , long term drinking of Pu-erh tea may be beneficial for the aged population in terms of increasing the body's resistance to infection and cancer . OUTPUT: avoiding immune destruction INPUT: Abstract The use of immunotherapeutics in melanoma has received much attention , and recent advances to further characterize the regulatory components of the immune system and the importance of co-stimulatory molecules have opened a new area for clinical investigation . Cytotoxic T lymphocyte-associated antigen 4 ( CTLA-4 ) serves as a negative regulator of immunity . Recent trials administering fully human anti-CTLA-4 monoclonal antibodies to melanoma patients have demonstrated clinically meaningful responses . Treatment with CTLA-4 blocking antibodies , however , is not without potential toxicities . Autoimmune side-effects , the most common being colitis-associated diarrhea , are frequently associated with clinical responses . In efforts to build upon prior vaccination efforts as well as attempt to offer patients clinically meaningful immune responses with a CTLA-4 blockade but without significant toxicities , we conducted a clinical trial in patients who previously received autologous tumor cells engineered to secrete granulocyte-macrophage colony stimulating factor ( GVAX ; Cell Genesys , South San Francisco , CA , USA ) with periodic infusions of CTLA-4 blocking antibodies . This sequential treatment resulted in clinically significant anti-tumor immunity without grade 3 or 4 toxicity in most patients . Pathological analyses following treatment of pre-existing tumors revealed a linear correlation between tumor necrosis and the ratio of intra-tumoral CD8+ effector cells to FoxP3+ regulatory cells ( T(regs) ) . Effective anti-tumor immunity and serious autoimmunity can be disassociated . Further targeting of anti-tumor T(regs)in combinatorial therapy approaches may be a rich avenue of future investigation . OUTPUT: avoiding immune destruction;resisting cell death INPUT: Interactions of CD70 , a tumor necrosis factor-related cell surface ligand and its receptor , CD27 , are thought to play an important role for T- , B- , and natural killer-cell activation . However , ligation of CD27 can also induce apoptosis . Human glioblastoma is paradigmatic for cancer-associated immunosuppression . We identified CD70 as a radioinducible gene in U87 MG glioma cells . A screening of a panel of human glioma cell lines revealed that 11 of 12 cell lines expressed CD70 mRNA and protein . Two human neuroblastoma cell lines did not express CD70 . CD70 mRNA expression was enhanced by irradiation in 8 of 12 glioma cell lines in a p53-independent manner . No alteration in CD70 expression was observed after glioma cell exposure to cytotoxic drugs such as lomustine . CD70 protein was also detected by immunocytochemistry in 5 of 12 glioblastomas and 3 of 4 anaplastic astrocytomas in vivo . CD27 expression was not detected in any glioma cell line , and there was no evidence for autocrine or backward signaling of the CD70 system in human glioma cells . Unexpectedly , CD70 expressed on glioma cells did not increase the immunogenicity of glioma cells in vitro . In contrast , CD70-positive glioma cells induced apoptosis in peripheral blood mononuclear cells ( PBMCs ) in a CD70-dependent manner . Neutralization of CD70 expressed on glioma cells prevented apoptosis and enhanced the release of tumor necrosis factor-alpha in cocultures of glioma cells and PBMCs . The effects of CD70-expressing glioma cells on PBMCs were mimicked by agonistic CD27 antibodies . Conversely , the shedding of CD27 by PBMCs was identified as a possible escape mechanism from glioma cell-induced CD70-dependent apoptosis . Thus , induction of B-cell and T-cell apoptosis via interactions of CD70 expressed on glioma cells and CD27 expressed on B and T cells may be a novel way for the immune escape of malignant gliomas . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Immunosuppression of immunoglobulin synthesis seen in patients with multiple myeloma is in part due to immunosuppressive CD5 positive B cells . In a 13 year longitudinal study of an IgA-deficient blood donor who developed multiple myeloma , the presence of immunosuppressive CD5 positive B cells and T cells preceded the diagnosis of overt multiple myeloma and the appearance of immunosuppressive monocytes . These data argue that certain immune defects may be involved in the development of myeloma and are not simply a consequence of overt malignancy . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot65

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Colorectal cancer ( CRC ) arises as the consequence of progressive changes from normal epithelial cells through polyp to tumor , and thus is an useful model for studying metabolic shift . In the present study , we studied the metabolomic profiles using high analyte specific gas chromatography/mass spectrometry ( GC/MS ) and liquid chromatography tandem mass spectrometry ( LC/MS/MS ) to attain a systems-level view of the shift in metabolism in cells progressing along the path to CRC . Colonic tissues including tumor , polyps and adjacent matched normal mucosa from 26 patients with sporadic CRC from freshly isolated resections were used for this study . The metabolic profiles were obtained using GC/MS and LC/MS/MS . Our data suggest there was a distinct profile change of a wide range of metabolites from mucosa to tumor tissues . Various amino acids and lipids in the polyps and tumors were elevated , suggesting higher energy needs for increased cellular proliferation . In contrast , significant depletion of glucose and inositol in polyps revealed that glycolysis may be critical in early tumorigenesis . In addition , the accumulation of hypoxanthine and xanthine , and the decrease of uric acid concentration , suggest that the purine biosynthesis pathway could have been substituted by the salvage pathway in CRC . Further , there was a step-wise reduction of deoxycholic acid concentration from mucosa to tumors . It appears that to gain a growth advantage , cancer cells may adopt alternate metabolic pathways in tumorigenesis and this flexibility allows them to adapt and thrive in harsh environment . OUTPUT: cellular energetics INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: Earlier studies indicated that density-arrested cancer cells released an unidentified growth inhibitor whose secretion was prevented by overexpression of the lysosomal protease cathepsin D ( cath D ) . In this study , this growth inhibitor was purified by affinity chromatography and identified as the heat shock cognate 70 protein ( hsc70 ) based on its peptide microsequencing and specific antibody recognition . Among intracellular proteins , including other heat shock proteins , only constitutive hsc70 was secreted in response to the high-cell density . Moreover , hsc70 secretion from cancer cells was generated by serum deprivation , whereas its cellular concentration did not change . Prevention of Hsc70 secretion by cath D overexpression was associated with the formation of multilayer cell cultures , thus indicating a loss of contact inhibition . In addition , we showed that supplementing the culture medium with purified hsc70 inhibited cell proliferation in the nanomolar range . Conversely , removal of this extracellular hsc70 from the medium by either retention on ADP-agarose or competition at the Hsc70 binding site restored cell proliferation . Hsc70 appears active in human breast cancer cells and hypersecreted by direct cath D inhibition . These results suggest a new role of this secreted hsc70 chaperone in cell proliferation that might account for the higher tumor growth of cancer cells overexpressing cath D . OUTPUT: evading growth suppressors INPUT: BACKGROUND The most deadly form of cancer is not lung or colon , breast or prostate ; it is any cancer that has become metastatic . Mortality due to metastatic melanoma , one of the most aggressive and deadly cancers , has increased steadily over the last several decades . Unfortunately , the arsenal of chemotherapeutic agents available today is most often unsuccessful at extending and improving the life expectancy of afflicted individuals . We sought to identify an effective and nontoxic agent against metastatic melanoma . METHODOLOGY/PRINCIPAL FINDINGS We chose to study Cloudman S-91 mouse melanoma cells ( sub-clone M3 , CCL53.1 ) because these cells are highly aggressive and metastatic , representing one of the deadliest types of cancer . Melanoma cells also had an experimental advantage because their morphology , which is easily monitored , relates to the physiology of metastatic cells and normal melanocytes . We chose to test methyl sulfone as a chemotherapeutic agent for two reasons . Because of its chemical structure , we speculated a potential anti-cancer activity by targeting microtubules . Equally important , methyl sulfone has a well-established safety profile in humans . Surprisingly , we found that malignant melanoma cells exposed to methyl sulfone demonstrated the loss of phenotypes characteristic of malignant cells , and the reemergence of phenotypes characteristic of healthy melanocytes . Briefly , over time methyl sulfone induced contact inhibition , loss of ability to migrate through an extracellular matrix , loss of anchorage-independent growth , proper wound healing followed by contact inhibition , irreversible senescence followed by arborization with melanosomes in arbors as seen in normal melanocytes . CONCLUSIONS/SIGNIFICANCE Methyl sulfone may have clinical potential as a non-toxic agent effective against metastatic melanoma . Additionally , methyl sulfone has promise as a tool to explore molecular mechanisms of metastatic transformation as well as fundamental processes such as cell migration , contact inhibition , wound healing and cellular senescence . OUTPUT: enabling replicative immortality;evading growth suppressors;activating invasion and metastasis INPUT: The influence of cell shape on the expression of proto-oncogenes was examined in normal and malignant human cells that varied in their sensitivities to contact-inhibition of proliferation . Cells were constrained into varying degrees of roundness by plating onto culture surfaces coated with different concentrations of poly(2-hydroxyethyl methacrylate ) ( poly[HEMA] ) and assayed for proliferation capacity and levels of c-myc , c-ras , c-fos , and c-fes mRNAs . Proliferation of contact-inhibited normal CUA-1 fibroblasts and the variant HT-IFNr cells was highly coupled to cell shape . As these cells became more rounded , a critical degree of roundness was reached at which proliferation ceased . In contrast , proliferation of non-contact-inhibited malignant HT-1080 cells was independent of cell shape . Northern analysis revealed that expression of c-myc and c-ras was highly sensitive to cell shape in the normal CUA-1 cells but not in the malignant HT-1080 or variant HT-IFNr cells . Levels of c-myc and c-ras mRNAs declined to nearly undetectable levels in CUA-1 cells at degrees of roundness that correlated with loss of proliferative ability . Expression of c-fos and c-fes oncogenes were independent of cell shape in all cells tested . Quantification of transcription rates by the nuclear run-off assay showed that shape modulation of c-myc and c-ras oncogene expression occurred at the transcriptional level . These data suggest that changes in cell shape can modulate expression of certain oncogenes and that these changes correlate with the cell's ability to proliferate . Moreover , inability to regulate c-myc and c-ras oncogene expression is associated with loss of shape-dependent growth controls and contact inhibition but that loss of this regulation alone is not sufficient to release cells from contact-inhibited controls . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot66

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Novel photosensitizers Hypocrellin A ( HA ) and Hypocrellin B ( HB ) , lipid soluble perylquinone derivatives of the genus Hypericum have a strong photodynamic effect on tumors and viruses . However , the mechanisms of tumor cell death induced by HA and HB are still unclear . In this study , we attempt to elucidate the photodynamic effects of HA and HB compounds in poorly differentiated ( CNE2 ) and moderately differentiated ( TW0-1 ) human nasopharyngeal carcinoma ( NPC ) cells as well as human mucosal colon ( CCL-220.1 ) and bladder ( SD ) cells . Using these cell lines we investigated few hall marks of apoptotic commitments in a drug and light dose dependent manner . Tumor cells photoactivated with HA and HB showed cell size shrinkage and an increase in the sub-diploid DNA content . A loss of membrane phospholipid asymmetry associated with apoptosis was induced by all tumor cell lines as evidenced by the externalization of phosphatidylserine . Western blot analysis of poly ( ADP-ribose ) polymerase , a caspases substrate , showed the classical cleavage pattern ( 116 to 85kDa ) associated with apoptosis in HA and HB-treated cell lysates . In addition , PARP cleavage was blocked by using tetrapepdide caspases inhibitors such as DEVD or z-VAD . These results demonstrate that tumor cell death induced by HB and HA is mediated by caspase proteases . This study also identifies both colon and bladder cells were more sensitive cell lines than NPC ( CNE2 and TWO-1 ) cell lines . OUTPUT: resisting cell death INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: We have previously demonstrated the antiproliferative effect of two flavonoids-2,2'-dihydroxychalcone ( DHC ) , a novel synthetic flavonoid , and fisetin , a naturally occurring flavonol-in prostate cancer cells . In this study , we further examine the mechanisms of these compounds on survival and proliferation pathways . DHC and fisetin ( 1-50 microM ) caused a dose-dependent reduction in viability , a concomitant increase in apoptosis in PC3 cells at 72 h , and a decrease in clonogenic survival at 24 h treatment . DHC was considerably more potent than fisetin in these cytotoxicity assays . The mechanism of accelerated cellular senescence was not activated by either compound in PC3 or lymph node carcinoma of the prostate ( LNCaP ) cells . Gene expression alterations in PC3 and LNCaP cells treated with 15 muM DHC and 25 microM fisetin for 6 to 24 h were determined by oligonucleotide array . Amongst the most highly represented functional categories of genes altered by both compounds was the cell cycle category . In total , 100 cell cycle genes were altered by DHC and fisetin including 27 genes with key functions in G2/M phase that were downregulated by both compounds . Other functional categories altered included chromosome organization , apoptosis , and stress response . These results demonstrate the multiple mechanisms of antitumor activity of DHC and fisetin in prostate cancer cells in vitro . OUTPUT: enabling replicative immortality;evading growth suppressors;sustaining proliferative signaling INPUT: Helicobacter hepaticus-infected Rag2(-/-) mice emulate many aspects of human inflammatory bowel disease , including the development of colitis and colon cancer . To elucidate mechanisms of inflammation-induced carcinogenesis , we undertook a comprehensive analysis of histopathology , molecular damage , and gene expression changes during disease progression in these mice . Infected mice developed severe colitis and hepatitis by 10wk post-infection , progressing into colon carcinoma by 20wk post-infection , with pronounced pathology in the cecum and proximal colon marked by infiltration of neutrophils and macrophages . Transcriptional profiling revealed decreased expression of DNA repair and oxidative stress response genes in colon , but not in liver . Mass spectrometric analysis revealed higher levels of DNA and RNA damage products in liver compared to colon and infection-induced increases in 5-chlorocytosine in DNA and RNA and hypoxanthine in DNA . Paradoxically , infection was associated with decreased levels of DNA etheno adducts . Levels of nucleic acid damage from the same chemical class were strongly correlated in both liver and colon . The results support a model of inflammation-mediated carcinogenesis involving infiltration of phagocytes and generation of reactive species that cause local molecular damage leading to cell dysfunction , mutation , and cell death . There are strong correlations among histopathology , phagocyte infiltration , and damage chemistry that suggest a major role for neutrophils in inflammation-associated cancer progression . Further , paradoxical changes in nucleic acid damage were observed in tissue- and chemistry-specific patterns . The results also reveal features of cell stress response that point to microbial pathophysiology and mechanisms of cell senescence as important mechanistic links to cancer . OUTPUT: tumor promoting inflammation;genomic instability and mutation;resisting cell death INPUT: Cell division and apoptosis are two crucial components of tumor biology and the importance of increased cell proliferation and reduced cell death have made them valid therapeutic targets . The plant kingdom is a relatively underexploited cache of novel drugs , and crude extracts of plants are known for their synergistic activity . The present study assessed the anti-proliferative activity of the medicinal plant Centrosema pubescens Benth . Centrosema pubescens dichloromethane extract ( CPDE ) inhibited the proliferation of HL-60 ( promyelocytic acute leukaemia ) cells with an IC₅₀ value of 5 μg/ml . Further studies also showed that CPDE induces growth arrest at the G1 phase and specifically down-regulates the expressions of cyclin E and CDK2 and up-regulates p27(CKI) levels . These events apparently lead to the induction of apoptosis , which was demonstrated qualitatively by a DNA fragmentation assay and propidium iodide staining . Quantitative assessment of the effective arrest of the cell cycle and of apoptosis was confirmed by flow cytometry . CPDE exhibited negligible cytotoxicity even at the highest dose tested ( 100 μg/ml ) in both normal peripheral blood mononuclear cells and in an in vitro model ( HL-60 ) . Our results strongly suggest that CPDE arrests the cell cycle at the G1 phase and triggers apoptosis by caspase activation . OUTPUT: evading growth suppressors;resisting cell death INPUT: Adhesion molecules play an important role in the functioning of the immune system , particularly with regard to cell-cell interactions and antigen presentation . Several adhesion molecules are expressed on Hodgkin's disease-derived cell lines and these are important in their molecular interactions as antigen presenting cells ( APC ) . There are no data regarding the expression of many of these adhesion molecules on Reed-Sternberg cells and its mononuclear variant ( Hodgkin's cells ( HC) ) present in pathological material . To obtain this information we undertook an immunohistological study on material from 18 cases of Hodgkin's disease using a panel of MoAbs to examine the expression of adhesion molecules on HC . The HC were shown to express the integrin beta 1 subfamily molecules , LFA-1 ( CD11a ) and p150,95 ( CD11c ) in high density but lacked CR3 ( CD11b ) . All of the immunoglobulin gene superfamily adhesion molecules studied were present to some degree on HC , with ICAM-2 , in particular , showing moderate to strong expression in most cases . The Hermes antigen CD44 was present in high density but leukosialin ( CD43 ) , another molecule present on diverse leucocyte types , was , in general , not detected on HC . These new data showing that ICAM-1 , ICAM-2 and LFA-3 are , like LFA-1 , expressed on HC emphasize the ability of HC to act as APC . The known adhesion molecule phenotype of the recently defined haematopoietic lineage of human dendritic cells ( DC ) is broadly similar to that of HC , perhaps supporting the hypothesis that some HC represent a malignancy of an APC ( DC ) lineage . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot67

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: DNA-protein cross-links ( DPCs ) are formed upon exposure to a variety of chemical and physical agents and pose a threat to genomic integrity . In particular , acrolein and related aldehydes produce DPCs , although the chemical linkages for such cross-links have not been identified . Here , we report that oligodeoxynucleotides containing 1,N(2)-deoxyguanosine adducts of acrolein , crotonaldehyde , and trans-4-hydroxynonenal can form cross-links with the tetrapeptide Lys-Trp-Lys-Lys . We concluded that complex formation is mediated by a Schiff base linkage because DNA-peptide complexes were covalently trapped following reduction with sodium cyanoborohydride , and pre-reduction of adducted DNAs inhibited complex formation . A previous NMR study demonstrated that duplex DNA catalyzes ring opening for the acrolein-derived gamma-hydroxy-1,N(2)-propanodeoxyguanosine adduct to yield an aldehydic function ( de los Santos , C. , Zaliznyak , T. , and Johnson , F . ( 2001 ) J. Biol . Chem. 276 , 9077-9082 ) . Consistent with this earlier observation , the adducts under investigation were more reactive in duplex DNA than in single-stranded DNA , and we concluded that the ring-open aldehydic moiety is the induced tautomer in duplex DNA for adducts exhibiting high relative reactivity . Adducted DNA cross-linked to Arg-Trp-Arg-Arg and Lys-Trp-Lys-Lys with comparable efficiency , and N(alpha)-acetylation of peptides dramatically inhibited trapping ; thus , the reactive nucleophile is located at the N-terminal alpha-amine of the peptide . These data suggest that Schiff base chemistry can mediate DPC formation in vivo following the formation of stable aldehyde-derived DNA adducts . OUTPUT: genomic instability and mutation INPUT: Often the use of cytotoxic drugs in cancer therapy results in stable disease rather than regression of the tumor , and this is typically seen as a failure of treatment . We now show that DNA damage is able to induce senescence in tumor cells expressing wild-type p53 . We also show that cytotoxics are capable of inducing senescence in tumor tissue in vivo . Our results suggest that p53 and p21 play a central role in the onset of senescence , whereas p16(INK4a) function may be involved in maintaining senescence . Thus , like apoptosis , senescence appears to be a p53-induced cellular response to DNA damage and an important factor in determining treatment outcome . OUTPUT: enabling replicative immortality;genomic instability and mutation;resisting cell death INPUT: Cis-diamminedichloroplatinum ( II ) ( cisplatin ) is a well characterized antitumor drug used for the treatment of a variety of human cancers . The cytotoxicity of cisplatin is mainly mediated through the formation of DNA adducts , which are also believed to be responsible for the secondary malignancies produced by the drug . The aim of this study was to determine the in vivo mutagenic activity of cisplatin in the lacZ plasmid-based transgenic mouse model . The mutant frequency ( MF ) and the spectrum of mutations induced by cisplatin in the mouse liver were analyzed and compared to controls . The mean MF in the lacZ gene was increased 2-fold in mice treated with a single 6 mg/kg body weight dose of cisplatin and sacrificed after 17 and 28 days ( P = 0.001 and P &lt ; 0.0001 ) . Restriction analysis and sequencing of mutant DNA showed that cisplatin was able to induce both large deletions and point mutations . A specific profile of base substitution and frameshift mutations was identified in treated mice , consisting primarily of G:C-->A:T transitions at GpG and ApG sites , the preferential DNA binding sites of cisplatin , and single basepair deletions/insertions . The present results provide the first evidence that cisplatin has mutagenic activity in vivo and induces a characteristic pattern of mutations in the mouse liver . This mutagenicity may be responsible for its tumorigenic activity . OUTPUT: genomic instability and mutation INPUT: The topoisomerase II ( topo II ) DNA incision-and-ligation cycle can be poisoned ( for example following treatment with cancer chemotherapeutics ) to generate cytotoxic DNA double-strand breaks ( DSBs ) with topo II covalently conjugated to DNA . Tyrosyl-DNA phosphodiesterase 2 ( Tdp2 ) protects genomic integrity by reversing 5'-phosphotyrosyl-linked topo II-DNA adducts . Here , X-ray structures of mouse Tdp2-DNA complexes reveal that Tdp2 β-2-helix-β DNA damage-binding ' grasp ' , helical ' cap ' and DNA lesion-binding elements fuse to form an elongated protein-DNA conjugate substrate-interaction groove . The Tdp2 DNA-binding surface is highly tailored for engagement of 5'-adducted single-stranded DNA ends and restricts nonspecific endonucleolytic or exonucleolytic processing . Structural , mutational and functional analyses support a single-metal ion catalytic mechanism for the exonuclease-endonuclease-phosphatase ( EEP ) nuclease superfamily and establish a molecular framework for targeted small-molecule blockade of Tdp2-mediated resistance to anticancer topoisomerase drugs . OUTPUT: genomic instability and mutation INPUT: In the course of a medicinal chemistry program aimed at discovering novel tumour-active rebeccamycin derivatives targeting DNA and/or topoisomerase I , a series of analogues with the sugar residue linked to the two indole nitrogens was recently developed . Two promising drug candidates in this staurosporine-rebeccamycin hybrid series were selected for a DNA-binding study reported here . The DNA interaction of the cationic indolocarbazole glycosides MP059 bearing a N,N-diethylaminoethyl side chain and MP072 containing a sugar bearing an amino group was compared with that of the uncharged analogue MP024 . The results show that the addition of a cationic substituent , either directly on the indolocarbazole chromophore or on the carbohydrate residue , significantly reinforces the interaction of the drugs with nucleic acids . The two cationic molecules MP059 and MP072 recognise preferentially sequences containing GpT.ApC and TpG.CpA steps but they do not inhibit topoisomerase I , in contrast to the parent uncharged derivative MP024 which stimulates DNA single strand breaks by topoisomerase I. The cytotoxic activity of the indolocarbazole derivatives bearing positively charged groups is one order of magnitude higher than that of the neutral compound MP024 . The high cytotoxic potential can be attributed to the enhanced DNA binding and sequence recognition capacity of the cationic compounds . The study provides useful information for further structure-activity relationship studies in the indolocarbazole series . OUTPUT: genomic instability and mutation INPUT: The chemotherapeutic drug cis-diamminedichloroplatinum ( II ) covalently binds to DNA resulting in a variety of adducts and cross-links which are thought to be responsible for the toxicity of the drug . We have used the gel mobility shift assay to detect proteins which bind to DNA treated in vitro with cis-diamminedichloroplatinum ( II ) and have identified two complexes which bind with increased affinity to cis-diamminedichloroplatinum ( II)-damaged DNA . Using monoclonal antibodies we have shown that one complex , B1 , contains human single-stranded DNA binding protein , a protein known to be involved in the in vitro repair synthesis assay of mammalian excision repair . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot68

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: When the prostate cancer cells become unresponsive to androgen therapy , resistance to chemotherapy becomes imminent , resulting in high mortality . To combat this situation , lycopodine , a pharmacologically important bioactive component derived from Lycopodium clavatum spores , was tested against hormone sensitive ( LnCaP ) and refractory ( PC3 ) prostate cancer cells in vitro . This study aims to check if lycopodine has demonstrable anti-cancer effects and if it has , to find out the possible mechanism of its action . The MTT assay was performed to evaluate the cytotoxic effect . Depolarization of mitochondrial membrane potential , cell cycle , EGF receptor activity and apoptosis were recorded by FACS ; profiles of different anti- and pro-apoptotic genes and their products were studied by semi-quantitative RT-PCR , indirect-ELISA , western blotting . Drug-DNA interaction was determined by CD spectroscopy . Administration of lycopodine down-regulated the expression of 5-lipoxygenase and the 5-oxo-ETE receptor ( OXE receptor1 ) and EGF receptor , and caused up-regulation of cytochrome c with depolarization of mitochondrial inner membrane potential , without palpable change in p53 activity , resulting in apoptosis , cell arrest at G0/G1 stage and ultimately reduced proliferation of cancer cells ; concomitantly , there was externalization of phosphotidyl serine residues . CD spectroscopic analysis revealed intercalating property of lycopodine with DNA molecule , implicating its ability to block cellular DNA synthesis . The overall results suggest that lycopodine is a promising candidate suitable for therapeutic use as an anti-cancer drug . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: Although the immense efforts have been made for cancer prevention , early diagnosis , and treatment , cancer morbidity and mortality has not been decreased during last forty years . Especially , lung cancer is top-ranked in cancer-associated human death . Therefore , effective strategy is strongly required for the management of lung cancer . In the present study , we found that novel daphnane diterpenoids , yuanhualine ( YL ) , yuanhuahine ( YH ) and yuanhuagine ( YG ) isolated from the flower of Daphne genkwa ( Thymelaeaceae ) , exhibited potent anti-proliferative activities against human lung A549 cells with the IC50 values of 7.0 , 15.2 and 24.7 nM , respectively . Flow cytometric analysis revealed that the daphnane diterpenoids induced cell-cycle arrest in the G0/G1 as well as G2/M phase in A549 cells . The cell-cycle arrests were well correlated with the expression of checkpoint proteins including the up-regulation of cyclin-dependent kinase inhibitor p21 and p53 and down-regulation of cyclin A , cyclin B1 , cyclin E , cyclin dependent kinase 4 , cdc2 , phosphorylation of Rb and cMyc expression . In the analysis of signal transduction molecules , the daphnane diterpenoids suppressed the activation of Akt , STAT3 and Src in human lung cancer cells . The daphnane diterpenoids also exerted the potent anti-proliferative activity against anticancer-drug resistant cancer cells including gemcitabine-resistant A549 , gefitinib- , erlotinib-resistant H292 cells . Synergistic effects in the growth inhibition were also observed when yuanhualine was combined with gemcitabine , gefitinib or erlotinib in A549 cells . Taken together , these findings suggest that the novel daphnane diterpenoids might provide lead candidates for the development of therapeutic agents for human lung cancers . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Tumor necrosis factor related apoptosis-inducing ligand ( TRAIL ) and agonistic anti-DR4/TRAIL-R1 and anti-DR5/TRAIL-R2 antibodies are currently under clinical investigation for treatment of different malignancies . TRAIL activates DR4 and DR5 and thereby triggers apoptotic and non-apoptotic signaling pathways , but possible different roles of DR4 or DR5 in these responses has poorly been addressed so far . In the present work , we analyzed cell viability , DISC formation as well as IL-8 and NF-kappaB activation side by side in responses to TRAIL and agonistic antibodies against DR4 ( mapatumumab ) and against DR5 ( lexatumumab ) in pancreatic ductal adenocarcinoma cells . We found that all three reagents are able to activate cell death and pro-inflammatory signaling . Death-inducing signaling complex ( DISC ) analysis revealed that mapatumumab and lexatumumab induce formation of homocomplexes of either DR4 or DR5 , whereas TRAIL additionally stimulated the formation of heterocomplexes of both receptors . Notably , blocking of receptors using DR4- and DR5-specific Fab fragments indicated that TRAIL exerted its function predominantly via DR4 . Interestingly , inhibition of PKC by Goe6983 enabled DR5 to trigger apoptotic signaling in response to TRAIL and also strongly enhanced lexatumumab-mediated cell death . Our results suggest the existence of mechanisms that silence DR5 for TRAIL- but not for agonistic-antibody treatment . OUTPUT: resisting cell death;tumor promoting inflammation INPUT: BACKGROUND Elevated levels of air pollution are associated with increased risk of lung cancer . Particulate matter ( PM ) contains transition metals that may potentiate neoplastic development through the induction of oxidative stress and inflammation , a lung cancer risk factor . Vanadium pentoxide ( V2O5 ) is a component of PM derived from fuel combustion as well as a source of occupational exposure in humans . In the current investigation we examined the influence of genetic background on susceptibility to V2O5-induced inflammation and evaluated whether V2O5 functions as a tumor promoter using a 2-stage ( initiation-promotion ) model of pulmonary neoplasia in mice . RESULTS A/J , BALB/cJ ( BALB ) , and C57BL/6J ( B6 ) mice were treated either with the initiator 3-methylcholanthrene ( MCA ; 10 microg/g ; i.p. ) or corn oil followed by 5 weekly aspirations of V2O5 or PBS and pulmonary tumors were enumerated 20 weeks following MCA treatment . Susceptibility to V2O5-induced pulmonary inflammation was assessed in bronchoalveolar lavage fluid ( BALF ) , and chemokines , transcription factor activity , and MAPK signaling were quantified in lung homogenates . We found that treatment of animals with MCA followed by V2O5 promoted lung tumors in both A/J ( 10.3 +/- 0.9 tumors/mouse ) and BALB ( 2.2 +/- 0.36 ) mice significantly above that observed with MCA/PBS or V2O5 alone ( P &lt ; 0.05 ) . No tumors were observed in the B6 mice in any of the experimental groups . Mice sensitive to tumor promotion by V2O5 were also found to be more susceptible to V2O5-induced pulmonary inflammation and hyperpermeability ( A/J>BALB>B6 ) . Differential strain responses in inflammation were positively associated with elevated levels of the chemokines KC and MCP-1 , higher NFkappaB and c-Fos binding activity , as well as sustained ERK1/2 activation in lung tissue . CONCLUSIONS In this study we demonstrate that V2O5 , an occupational and environmentally relevant metal oxide , functions as an in vivo lung tumor promoter among different inbred strains of mice . Further , we identified a positive relationship between tumor promotion and susceptibility to V2O5-induced pulmonary inflammation . These findings suggest that repeated exposures to V2O5 containing particles may augment lung carcinogenesis in susceptible individuals through oxidative stress mediated pathways . OUTPUT: tumor promoting inflammation INPUT: To elucidate the mechanism of the synergistic cytotoxicity of 5-fluorouracil ( 5-FU ) and cis-diamminedichloroplatinum(II) ( CDDP ) , we studied the interaction of these agents using a human squamous carcinoma cell line ( HST-1 ) . Exposure to 5-FU for 24 h and to CDDP for 1 h produced a 50% inhibitory concentration of 1.0 micrograms/ml ( 7.7 microM ) and 2.5 micrograms/ml ( 8.3 microM ) , respectively . The cytotoxic action of CDDP was augmented , and a greater than additive effect was observed when the cells were exposed to 5-FU ( 1.0 micrograms/ml ; 7.7 microM ) for 24 h before the CDDP treatment . This synergistic activity was maximal when the interval between 5-FU and CDDP exceeded 24 h . In contrast , the cytotoxicity of CDDP was attenuated when it preceded the exposure to 5-FU . Thymidine did not alter the 5-FU-CDDP interaction . Evaluation of the kinetics of the removal of DNA interstrand cross-links , measured by alkaline elution , showed a significant reduction of this removal in the cells exposed to 5-FU followed by CDDP with a drug-free interval of 48 h , as compared with cells exposed to CDDP alone , or to 5-FU immediately followed by CDDP , although no differences were found in the formation of DNA interstrand cross-links by CDDP among these cells . No significant differences in the accumulation of intracellular platinum were detected by atomic absorption spectrophotometry . These findings suggest that 5-FU modulates the repair of platinum-DNA adducts , thereby potentiating the antitumor activity of CDDP . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot69

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Metastasis is a major cause of death of patients with malignant tumors . Matrix metalloproteinases ( MMPs ) are important for the migration and invasion of various types of cancer cell . Propofol is a known anesthetic agent , widely used for short-term anesthesia and for longer-term sedation . Propofol inhibits the proliferation of a variety of tumor cells , but there is no available information regarding propofol-inhibited migration and invasion of tumor cells in vitro . In this study , we investigated the effects of propofol on the migration and invasion of human lung carcinoma A549 cells . Wound healing assay and Boyden chamber assays indicated that propofol inhibited the migration and invasion of A549 cells in vitro . Gelatin zymographic analysis showed the inhibitory effect of propofol on the activation of expression MMP-2 . Western blot analysis also indicated that propofol suppressed the protein expiration of growth factor receptor-bound protein 2 ( GRB2 ) , Jun N-terminal kinases 1/2 ( p-JNK1/2 ) , p-p38 , MMP-2 and MMP-9 in A549 cells . Results from real-time PCR assay also showed that propofol inhibited the mRNA gene expression of MMP-2 , -7 and -9 , and enhanced that of tissue inhibitor of metalloproteinase 1 ( TIMP1 ) and TIMP2 in A549 cells . Taken together , these data show that propofol inhibits MMP-2 and -9 mRNA and protein expressions , resulting in suppression of lung cancer cell invasion and migration in vitro . OUTPUT: activating invasion and metastasis INPUT: OBJECTIVE To observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe ( RSR ) on HER2 positive breast cancer , to evaluate the effects of RSR on the expressions of matrix metalloproteinases ( MMPs ) and the tissue inhibitor of metalloproteinases ( TIMPs ) in the recurrence and metastasis of HER2 positive breast cancer , thus revealing its anti-tumor recurrent and metastatic mechanisms . METHODS Selected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu . The primary tumor resection was carried out . After surgery they were randomly divided into the blank control group , the RSR group , the Herceptin group , and the combination group ( RSR + Herceptin group ) . The treatment lasted for 4 months . The inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated . The expressions of matrix metalloproteinase-2 ( MMP-2 ) , matrix metalloproteinase-9 ( MMP-9 ) , tissue inhibitor of metalloproteinase ( TIMP-1 ) , and TIMP-2 in the recurrent tumor tissue were detected using Western blot . RESULTS By the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group , showing statistical difference between the two groups ( P = 0.037 ) . The average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group . The inhibition rate of lung metastasis was 37. 85% in the RSR group , but with no statistical significance . The expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group , the Herceptin group , and the combination group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group , the Herceptin group , and the combination group was significantly down-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group ( P &lt ; 0.05 ) . The expressions of both TIMP-1 and TIMP-2 of the RSR group , the Herceptin group , and the combination group were all up-regulated when compared with the blank control group ( P &lt ; 0.05 ) . The increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group ( P &lt ; 0.05 ) . It was higher in the combination group than in the RSR group ( P &lt ; 0.05 ) . CONCLUSIONS RSR could inhibit the tumor recurrence of FVB/neu mice . It could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix . It might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells . OUTPUT: activating invasion and metastasis INPUT: Numerous studies have reported a correlation between production of 72-kDa ( MMP-2 ) and 92-kDa ( MMP-9 ) type-IV collagenases/gelatinases and the metastatic potential of cancer cells . An abrogating effect of tissue inhibitors of metalloproteinases ( TIMP-1 and TIMP-2 ) on metastases has also been noted . In this report we have used sensitive enzyme-linked immunoassays to measure MMP-2 , MMP-9 , TIMP-1 and TIMP-2 levels in eight human lung-cancer cell lines which were characterized for biological behavior in nude mice . We demonstrated that the Calu-6 and A549 cell lines with the highest metastatic , invasive and tumorigenic potential secreted the highest levels of MMP-2 . MMP-9 and TIMP-1 secretions were comparatively low in all cell lines . TIMP-2 secretion , which exceeded MMP-2 secretion for all cell lines , did not correlate with metastatic potential . To further explore these correlations , the metastatic Calu-6 cell line was transfected with a K-rev-1 cDNA expression construct . The K-rev revertant cell lines demonstrated a more differentiated phenotype and were less tumorigenic , invasive and metastatic in nude mice . Nonetheless , the Calu-6 revertant cell lines secreted higher levels of MMP-2 than the parent cell line . In conclusion , invasion and metastasis by lung-cancer cells requires not only enhanced MMP production , but also other less well-understood tumorigenic characteristics . The multiplicity of factors required by cancer cells for dissemination helps to explain the minute fraction of cancer cells from a primary tumor that ever develop into a metastasis . OUTPUT: activating invasion and metastasis INPUT: Background and Objective . The cell cycle is regulated by proteins at different checkpoints , and dysregulation of this cycle plays a role in carcinogenesis . Matrix metalloproteinases ( MMPs ) are enzymes that degrade collagen and promote tumour infiltration . The aim of this study was to evaluate the expression of various cell cycle regulators and MMPs and to correlate such expression with progression and recurrence in patients with stage T1 urothelial carcinoma of the bladder ( UCB ) . Patients and Methods . This population-based cohort study comprised 201 well-characterized patients with primary stage T1 urothelial carcinoma of the bladder . Immunohistochemistry was performed on formalin-fixed material to quantify expression of cell cycle regulators and two MMPs . Results . Normal expression of p53 and abnormal expression of MMP9 were associated with greater risk of tumour recurrence . Also , normal p16 expression was related to a lower risk of tumour progression . MMP2 , p21 , cyclin D1 , and pRb showed no significant results that could estimate progression or recurrence . Conclusions . Normal p16 expression is associated with a lower risk of tumour progression , but immunohistochemistry on cell cycle regulators and MMPs has little value in predicting the prognosis in stage T1 UCB . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: High-grade prostate cancers express high levels of matrix metalloproteinases ( MMPs ) , major enzymes involved in tumor invasion and metastasis . However , the tumor cell lines commonly employed for prostate cancer research express only small amounts of MMPs when cultivated as monolayer cultures , in common culture media . The present study was conducted to ascertain whether culture conditions that include fibronectin can alter MMP2 and MMP9 expression by the human prostatic epithelial cell lines RWPE-1 , LNCaP and PC-3 . These cells were individually seeded at 2×10(4) cells/cm(2) , cultivated until they reached 80% confluence , and then exposed for 4h to fibronectin , after which the conditioned medium was analyzed by gelatin zymography . Untreated cells were given common medium . Only RWPE-1 cells express detectable amounts of MMP9 when cultivated in common medium , whereas the addition of fibronectin induced high expression levels of pro and active forms of MMP2 in all tested cell lines . Our findings demonstrate that normal and tumor prostate cell lines express MMP2 activity when in contact with extracellular matrix components or blood plasma proteins such as fibronectin . Future studies of transcriptomes and proteomes in prostate cancer research using these cell lines should not neglect these important conclusions . OUTPUT: activating invasion and metastasis INPUT: BACKGROUND Matrix metalloproteinases ( MMP ) are a gene family of zinc enzymes capable of degrading almost all of the extracellular matrix macromolecules in vivo . Their enzymic activities are believed to be responsible for tumor invasion and metastasis . METHODS In this study , using peroxidase-antiperoxidase method , monospecific antisera against MMP-1 ( tissue collagenase ) , MMP-2 ( type IV collagenase/72-kilodalton [ KD ] gelatinase ) , and MMP-3 ( stromelysin ) were applied to 29 squamous cell carcinomas and normal epithelium of the esophagus to identify cells synthesizing and secreting these enzymes . RESULTS Immunoreactivity of MMP-1 , -2 , and -3 was observed in small cancer nests of the deeply invasive or marginal portion of the tumor . Among the 29 patients studied , the presence of at least one MMP was observed in 17 ( 58.6% ) . All three enzymes were observed in six ( 20.6% ) patients , MMP-2 and -3 in five ( 17.2% ) patients , only MMP-2 in three ( 10.3% ) patients , and MMP-3 alone in three ( 10.3% ) patients . There was a good correlation among histologic stage and tumor invasion , lymph node metastasis , and MMP expression . In particular , expression of MMP-2 and -3 was closely related to lymph node metastasis and vascular invasion . CONCLUSIONS These results suggest that MMP , especially MMP-2 and -3 , play an important role in tumor invasion and metastasis and that analysis of MMP-2 and -3 production is useful for evaluation of malignant potential in esophageal carcinoma . OUTPUT:

activating invasion and metastasis

HoC_dynamic_5_shot70

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Blockage of the metastasis process remains a significant clinical challenge , requiring innovative therapeutic approaches . For this purpose , molecules that inhibit matrix metalloproteinases activity or induce the expression of their natural inhibitor , the tissue inhibitor of metalloproteinases ( TIMPs ) , are potentially interesting . In a previous study , we have shown that synthetic ligands binding to cell surface nucleolin/nucleophosmin and known as HB 19 for the lead compound and NucAnt 6L ( N6L ) for the most potent analog , inhibit both tumor growth and angiogenesis . Furthermore , they prevent metastasis in a RET transgenic mice model which develops melanoma . Here , we investigated the effect of N6L on the invasion capacity of MDA-MB-435 melanoma cells . Our results show that the multivalent pseudopeptide N6L inhibited Matrigel invasion of MDA-MB-435 cells in a modified Boyden chamber model . This was associated with an increase in TIMP-3 in the cell culture medium without a change in TIMP-3 mRNA expression suggesting its release from cell surface and/or extracellular matrix . This may be explained by our demonstrated N6L interaction with sulfated glycosaminoglycans and consequently the controlled bioavailability of glycosaminoglycan-bound TIMP-3 . The implication of TIMP-3 in N6L-induced inhibition of cell invasion was evidenced by siRNA silencing experiments showing that the loss of TIMP-3 expression abrogated the effect of N6L . The inhibition of tumor cell invasion by N6L demonstrated in this study , in addition to its previously established inhibitory effect on tumor growth and angiogenesis , suggests that N6L represents a promising anticancer drug candidate warranting further investigation . OUTPUT: activating invasion and metastasis;inducing angiogenesis INPUT: Increasing evidence shows that estrogens are involved in lung cancer proliferation and progression , and most human lung tumors express estrogen receptor β ( ERβ ) as well as aromatase . To determine if the aromatase inhibitor anastrozole prevents development of lung tumors induced by a tobacco carcinogen , alone or in combination with the ER antagonist fulvestrant , ovariectomized female mice received treatments with the tobacco carcinogen 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone ( NNK ) along with daily supplements of androstenedione , the substrate for aromatase . Placebo , anastrozole and/or fulvestrant were administered in both an initiation and a promotion protocol of lung tumorigenesis . The combination of fulvestrant and anastrozole given during NNK exposure resulted in significantly fewer NNK-induced lung tumors ( mean = 0.5 ) compared with placebo ( mean = 4.6 , P &lt ; 0.001 ) , fulvestrant alone ( mean = 3.4 , P &lt ; 0.001 ) or anastrozole alone ( mean = 2.8 , P = 0.002 ) . A significantly lower Ki67 cell proliferation index was also observed compared with single agent and control treatment groups . Beginning antiestrogen treatment after NNK exposure , when preneoplastic lesions had already formed , also yielded maximum antitumor effects with the combination . Aromatase expression was found mainly in macrophages infiltrating preneoplastic and tumorous areas of the lungs , whereas ERβ was found in both macrophages and tumor cells . Antiestrogens , especially in combination , effectively inhibited tobacco carcinogen-induced murine lung tumorigenesis and may have application for lung cancer prevention . An important source of estrogen synthesis may be inflammatory cells that infiltrate the lungs in response to carcinogens , beginning early in the carcinogenesis process . ERβ expressed by inflammatory and neoplastic epithelial cells in the lung may signal in response to local estrogen production . OUTPUT: sustaining proliferative signaling INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: INTRODUCTION Necrosis at the tumor center is a common feature of aggressive breast cancers and has been associated with poor prognosis . It is commonly identified by means of invasive histopathology , which often correlates with morbidity and potential tumor cell dissemination , and limits the reconstruction of the whole necrotic domain . In this study we hypothesized that non covalent association to serum albumin ( SA ) and covalent binding to ligands for tumor-abundant cell receptors should synergistically drive selective accumulation and prolonged retention of imaging and therapeutic agents in breast tumor necrotic domains enabling in vivo identification , imaging and possibly treatment of such tumors . METHODS Cyclo-Arg-Gly-Asp-D-Phe-Lys ( c(RGDfK) ) were conjugated to bacteriochlorophyll-derivatives ( Bchl-Ds ) , previously developed as photodynamic agents , fluorescent probes and metal chelators in our lab . The c(RGDfK) component drives ligation to alphaVbeta3 integrin receptors over-expressed by tumor cells and neo-vessels , and the Bchl-D component associates to SA in a non-covalent manner . STL-6014 , a c(RGDfK)-Bchl-D representative , was i.v. injected to CD-1 , nude female mice bearing necrotic and non-necrotic human MDA-MB-231-RFP breast cancer tumors . The fluorescence signals of the Bchl-Ds and RFP were monitored over days after treatment , by quantitative whole body imaging and excised tumor/tissue samples derived thereof . Complementary experiments included competitive inhibition of STL-6014 uptake by free c(RGDfK) , comparative pharmacokinetics of nonconjugated c(RGDfK) Bchl-D ( STL-7012 ) and of two human serum albumin ( HSA ) conjugates : HSA-STL-7012 and HSA-STL-6014 . RESULTS STL-6014 and STL-7012 formed complexes with HSA ( HSA/STL-6014 , HSA/STL-7012 ) . STL-6014 , HSA-STL-7012 and HSA-STL-6014 , selectively accumulated at similar rates , in tumor viable regions over the first 8 h post administration . They then migrated into the necrotic tumor domain and presented tumor half lifetimes ( T1/2 ) in the range of days where T1/2 for HSA-STL-6014 &gt ; STL-6014 &gt ; HSA-STL-7012 . No accumulation of STL-7012 was observed . Pre-injection of c(RGDfK) excess , prevented the uptake of STL-6014 in the small , but not in the large tumors . CONCLUSIONS Non-covalent association to SA and covalent binding to c(RGDfK) , synergistically enable the accumulation and prolonged retention of Bchl-Ds in the necrotic regions of tumors . These findings provide novel guidelines and strategy for imaging and treatment of necrotic tumors . OUTPUT: resisting cell death INPUT: The results of experimental studies have indicated the pleiotropic effects of statins in organism , e.g. the influence on cell cycle , apoptosis or angiogenesis . In this study , the effects of simvastatin on selected parameters of apoptosis and proliferation in chemocarcinogen-induced mammary tumorigenesis in female rats were determined . Simvastatin was administered dietary at a dose of 18 mg/kg and highly effective dose of 180 mg/kg the entire experiment ( 18 weeks ) . At autopsy mammary tumors were removed and prepared for immunohistochemical and histomorphological analysis . In treated animals ( simvastatin 180 mg/kg ) , significant decrease by 12% in Bcl-2 protein expression and non-significant decrease by 27% of Ki67 protein expression in tumor cells compared to tumor cells in control animals were observed after semiquantitative evaluation . Morphometrical analysis has shown significant proapototic shift in Bcl-2/Bax ratio in tumor cells . In high grade control carcinoma cells , the expression of Ki67 increased by 37% ( non-significantly ) in comparison with control low grade carcinomas . A histomorphological analysis of malignant tumors has revealed a shift from high grade to low grade carcinomas after simvastatin treatment . The noticeable decrease of mammary tumor frequency and incidence in rats after simvastatin treatment was accompanied with antiapoptotic Blc-2 protein decrease and proapoptotic Bax protein increase in this experiment . OUTPUT: resisting cell death INPUT: Nine primary pulmonary carcinomas , one metastatic carcinoma , and two malignant pleural mesotheliomas have been analysed for the expression at the mRNA level of metalloproteinases ( MPs ) and tissue inhibitors of MPs ( TIMPs ) . In situ hybridisation showed TIMP-1 and TIMP-2 transcripts predominantly over tumour stroma and gelatinases evenly distributed over both stromal and tumour cells . While both TIMP-1 and TIMP-2 were expressed in non-neoplastic lungs ( NNL ) as well as in carcinomas , stromelysin 3 ( ST3 ) , 92 kDa gelatinase and interstitial collagenase were expressed only by carcinomas . Expression of these MPs by carcinomas was independent of histologic type and such tumour features as fibrosis or necrosis . The consistent expression of ST3 by all of the carcinomas examined and absence of its expression in NNL indicates that ST3 production is likely associated with the malignant phenotype . However , since 92 kDa gelatinase and interstitial collagenase transcripts were found in some but not all tumour samples , their expression is not a uniform feature of pulmonary carcinomas . The possible prognostic significance of the expression of the latter two enzymes by carcinomas remains to be established . OUTPUT:

resisting cell death

HoC_dynamic_5_shot71

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: AIM To investigate whether luteolin , a highly prevalent flavonoid , reverses the effects of epithelial-mesenchymal transition ( EMT ) in vitro and in vivo and to determine the mechanisms underlying this reversal . METHODS Murine malignant melanoma B16F10 cells were exposed to 1% O(2) for 24 h . Cellular mobility and adhesion were assessed using Boyden chamber transwell assay and cell adhesion assay , respectively . EMT-related proteins , such as E-cadherin and N-cadherin , were examined using Western blotting . Female C57BL/6 mice ( 6 to 8 weeks old ) were injected with B16F10 cells ( 1�10(6) cells in 0.2 mL per mouse ) via the lateral tail vein . The mice were treated with luteolin ( 10 or 20 mg/kg , ip ) daily for 23 d . On the 23rd day after tumor injection , the mice were sacrificed , and the lungs were collected , and metastatic foci in the lung surfaces were photographed . Tissue sections were analyzed with immunohistochemistry and HE staining . RESULTS Hypoxia changed the morphology of B16F10 cells in vitro from the cobblestone-like to mesenchymal-like strips , which was accompanied by increased cellular adhesion and invasion . Luteolin ( 5-50 μmol/L ) suppressed the hypoxia-induced changes in the cells in a dose-dependent manner . Hypoxia significantly decreased the expression of E-cadherin while increased the expression of N-cadherin in the cells ( indicating the occurrence of EMT-like transformation ) , which was reversed by luteolin ( 5 μmol/L ) . In B16F10 cells , luteolin up-regulated E-cadherin at least partly via inhibiting the β3 integrin/FAK signal pathway . In experimental metastasis model mice , treatment with luteolin ( 10 or 20 mg/kg ) reduced metastatic colonization in the lungs by 50% . Furthermore , the treatment increased the expression of E-cadherin while reduced the expression of vimentin and β3 integrin in the tumor tissues . CONCLUSION Luteolin inhibits the hypoxia-induced EMT in malignant melanoma cells both in vitro and in vivo via the regulation of β3 integrin , suggesting that luteolin may be applied as a potential anticancer chemopreventative and chemotherapeutic agent . OUTPUT: activating invasion and metastasis INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Focal inflammation causes systemic fever . Cancer hyperthermia therapy results in shrinkage of tumors by various mechanisms , including induction of adaptive immune response . However , the physiological meaning of systemic fever and mechanisms of tumor shrinkage by hyperthermia have not been completely understood . In this study , we investigated how heat shock influences the adaptive immune system . We established a cytotoxic T lymphocyte ( CTL ) clone ( #IM29 ) specific for survivin , one of the tumor-associated antigens ( TAAs ) , from survivin peptide-immunized cancer patients ' peripheral blood , and the CTL activities were investigated in several temperature conditions ( 37-41�C ) . Cytotoxicity and IFN-γ secretion of CTL were greatest under 39�C condition , whereas they were minimum under 41�C . To address the molecular mechanisms of this phenomenon , we investigated the apoptosis status of CTLs , expression of CD3 , CD8 , and TCRαβ by flow cytometry , and expression of perforin , granzyme B , and Fas ligand by western blot analysis . The expression of perforin and granzyme B were upregulated under temperature conditions of 39 and 41�C . On the other hand , CTL cell death was induced under 41�C condition with highest Caspase-3 activity . Therefore , the greatest cytotoxicity activity at 39�C might depend on upregulation of cytotoxic granule proteins including perforin and granzyme B. These results suggest that heat shock enhances effector phase of the adaptive immune system and promotes eradication of microbe and tumor cells . OUTPUT: resisting cell death INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Fifty-six previously untreated stage-I ( according to Rai ) chronic lymphocytic leukemia ( CLL ) patients were examined for their clinical data , immunological characteristics , and hormonal values . Dysfunction of T and B lymphocytes was demonstrated by changed lymphocyte blastogenic response to stimulation with phytohemagglutinin ( PHA ) , concanavalin A ( ConA ) , pisum sativatum agglutinin ( PSA ) , wheat germ agglutinin ( WGA ) , recombinant interleukin 2 ( IL 2 ) , and dextran sulfate ( DxS ) ; also by decreased immunoglobulin levels ( IgG , IgA , IgE ) and increased beta 2-microglobulin ( beta 2-M ) values . Simultaneously , dysregulation of the hypothalamic-pituitary-adrenal axis , immune system integration , imbalance of sex hormones , and changes in thyroid hormones were observed in the same group of patients . Disturbed immunohormonal interactions in early-stage CLL may be responsible for the pathogenetic mechanisms in this lymphoproliferative malignancy . OUTPUT: avoiding immune destruction INPUT: Studies were performed to test the hypothesis that urethane-induced murine lung tumors exhibit xenobiotic resistance and alterations in pulmonary cytochrome P-450 enzymes. 1,1-Dichloroethylene , naphthalene , and paraquat were administered to tumor-bearing and control mice to elicit acute lung cytotoxicity , and responses were evaluated in tumors ( papillary and solid ) , uninvolved surrounding tissue , and untreated control lung. 1,1-Dichloroethylene ( 125 mg/kg , i.p. ) and naphthalene ( 225 mg/kg , i.p. ) caused preferential necrosis of Clara cells in control lungs and uninvolved tissue of tumor-bearing lungs . In contrast , papillary and solid tumors were both resistant to 1,1-dichloroethylene-induced cytotoxicity . Paraquat ( 10 , 20 mg/kg , i.v. ) elicited Clara cell damage in control lungs and uninvolved lung tissue of tumor-bearing mice , with minor disruption of the alveolar epithelium . Neither papillary nor solid tumors sustained any apparent cell damage from paraquat . Immunoblots of P-450 enzymes confirmed constitutive expression of CYP2B1 in control lung and uninvolved lung tissue of tumor-bearing mice , but this P-450 enzyme was not detected in either adenomas or carcinomas . Lung CYP1A1 was inducible by beta-naphthoflavone in non-tumor-bearing mice and uninvolved tissue of tumor-bearing mice ; however , inducibility was decreased in adenomas and abolished in carcinomas . These results demonstrate resistance of lung tumor cells to chemically induced cytotoxicity and diminished expression of cytochrome P-450 enzymes in tumors . OUTPUT:

resisting cell death

HoC_dynamic_5_shot72

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: INTRODUCTION Tamoxifen is the most widely prescribed anti-estrogen treatment for patients with estrogen receptor ( ER)-positive breast cancer . However , there is still a need for biomarkers that reliably predict endocrine sensitivity in breast cancers and these may well be expressed in a dynamic manner . METHODS In this study we assessed gene expression changes at multiple time points ( days 1 , 2 , 4 , 7 , 14 ) after tamoxifen treatment in the ER-positive ZR-75-1 xenograft model that displays significant changes in apoptosis , proliferation and angiogenesis within 2 days of therapy . RESULTS Hierarchical clustering identified six time-related gene expression patterns , which separated into three groups : two with early/transient responses , two with continuous/late responses and two with variable response patterns . The early/transient response represented reductions in many genes that are involved in cell cycle and proliferation ( e.g . BUB1B , CCNA2 , CDKN3 , MKI67 , UBE2C ) , whereas the continuous/late changed genes represented the more classical estrogen response genes ( e.g . TFF1 , TFF3 , IGFBP5 ) . Genes and the proteins they encode were confirmed to have similar temporal patterns of expression in vitro and in vivo and correlated with reduction in tumour volume in primary breast cancer . The profiles of genes that were most differentially expressed on days 2 , 4 and 7 following treatment were able to predict prognosis , whereas those most changed on days 1 and 14 were not , in four tamoxifen treated datasets representing a total of 404 patients . CONCLUSIONS Both early/transient/proliferation response genes and continuous/late/estrogen-response genes are able to predict prognosis of primary breast tumours in a dynamic manner . Temporal expression of therapy-response genes is clearly an important factor in characterising the response to endocrine therapy in breast tumours which has significant implications for the timing of biopsies in neoadjuvant biomarker studies . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: The published results on nanoparticles cytotoxicity and genotoxicity such as titanium dioxide nanoparticles ( TiO(2) NPs ) are inconsistent , and often conflicting and insufficient . Since different parameters may have impact on the toxicity results , there is need to lay stress on detailed characterization of NPs and the use of different testing conditions for assessment of NPs toxicity . In order to investigate whether dispersion procedures influence NP cytotoxicity and genotoxicity , we compared two protocols giving TiO(2) NP dispersions with different stability and agglomeration states . Detailed primary and secondary characteristics of both TiO(2) NP dispersions in culture media were carried out before toxicological testing ; TK6 human lymphoblast cells , EUE human embryonic epithelial cells and Cos-1 monkey kidney fibroblasts were used to assess cytotoxicity ( by trypan blue exclusion , proliferation activity and plating efficiency assays ) and genotoxicity ( by the comet assay ) . DNA strand breaks were detected by the alkaline comet assay . DNA oxidation lesions ( especially 8-oxo-7,8-dihydroguanine , 8-oxoG ) were measured with a modified comet assay including incubation with specific repair enzyme formamidopyrimidine DNA glycosylase ( FPG ) . The TiO(2) NPs dispersion with large agglomerates ( 3 min sonication and no serum in stock solution ) induced DNA damage in all three cell lines , while the TiO(2) NPs dispersed with agglomerates less than 200 nm ( foetal serum in stock solution and sonication 15 min ) had no effect on genotoxicity . An increased level of DNA oxidation lesions detected in Cos-1 and TK6 cells indicates that the leading mechanism by which TiO(2) NPs trigger genotoxicity is most likely oxidative stress . Our results show that the dispersion method used can influence the results of toxicity studies . Therefore at least two different dispersion procedures should be incorporated into assessment of cyto- and genotoxic effects of NPs . It is important , when assessing the hazard associated with NPs , to establish standard testing procedures and thorough strategies to consider the diverse conditions relevant to possible exposures . OUTPUT: genomic instability and mutation;tumor promoting inflammation INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: Twelve postmenopausal women ( 54-93 years ) with primary breast carcinoma were treated with tamoxifen due to infirmity or refusal to undergo surgery . Seven premenopausal patients ( 32-50 years ) were given preoperative chemotherapy because of large tumors or inflammatory carcinoma . Fine-needle aspiration biopsy was used to procure tumor cells for diagnosis , hormone receptor determination and analysis of proliferation fraction . Aspirations were repeated every 3 months in the tamoxifen group and each month in patients receiving chemotherapy . Two patients who responded to tamoxifen had tumors with more than 75% estrogen receptor positive cells . A decreased proliferation fraction was observed in two tumors responding to tamoxifen . Eight patients , all with estrogen receptor positive tumors , had stable disease . Progressive disease was observed in two patients with less than 25% receptor positive cells . In these tumors the percentage of proliferating cells remained high during therapy . Objective response was recorded for six patients treated with chemotherapy . The clinical response was reflected in a decreased proliferation fraction . No correlation was observed between response and percentage of proliferating cells in the untreated tumor . The results suggest that analysis of tumor cell characteristics such as hormone receptor content and proliferation fraction can be used to predict and monitor response to endocrine treatment and chemotherapy in breast carcinomas . OUTPUT: sustaining proliferative signaling INPUT: We studied the effects of double-strand breaks on intramolecular extrachromosomal homologous recombination in mammalian cells . Pairs of defective herpes thymidine kinase ( tk ) sequences were introduced into mouse Ltk- cells on a DNA molecule that also contained a neo gene under control of the SV40 early promoter/enhancer . With the majority of the constructs used , gene conversions or double crossovers , but not single crossovers , were recoverable . DNA was linearized with various restriction enzymes prior to transfection . Recombination events producing a functional tk gene were monitored by selecting for tk-positive colonies . For double-strand breaks placed outside of the region of homology , maximal recombination frequencies were measured when a break placed the two tk sequences downstream from the SV40 early promoter/enhancer . We observed no relationship between recombination frequency and either the distance between a break and the tk sequences or the distance between the tk sequences . The quantitative effects of the breaks appeared to depend on the degree of homology between the tk sequences . We also observed that inverted repeats recombined as efficiently as direct repeats . The data indicated that the breaks influenced recombination indirectly , perhaps by affecting the binding of a factor(s) to the SV40 promoter region which in turn stimulated or inhibited recombination of the tk sequences . Taken together , we believe that our results provide strong evidence for the existence of a pathway for extrachromosomal homologous recombination in mammalian cells that is distinct from single-strand annealing . We discuss the possibility that intrachromosomal and extrachromosomal recombination have mechanisms in common . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot73

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Spindle cell oncocytoma ( SCO ) of the pituitary gland is a relatively recently established , very rare subtype of adenohypophysis tumours that was introduced as a distinct clinicopathological entity in the fourth edition of WHO classification of the central nervous system tumours ( 2007 ) . It is non-endocrine neoplasm of the anterior pituitary that occurs in adults and usually follows a benign clinical course , corresponding to WHO grade I. Up to now , pituitary SCO have been reported occasionally and only 14 cases of SCO have been documented in the literature . Because of their rarity , the pathogenesis and natural history of these tumours have not been fully characterized . We report two additional cases of SCO occurring in females aged 63 years ( Case 1 ) and 65 years ( Case 2 ) , who presented with pan-hypopituitarism , headache and visual field defect . In both cases , the magnetic resonance imaging showed solid sellar mass of moderate size with suprasellar extension . The clinical and radiological features suggested non-functioning pituitary macroadenomas without evidence of invasive growth . One patient presented with tumour recurrence 3 years after undergoing the previous surgical removal of tumour , which was initially misdiagnosed as schwannoma . The first tumour was removed by transsphenoidal surgery and the second one by frontal craniotomy . Histologically and immunohistochemically , both tumours displayed the features typical for SCO of the pituitary . They were composed of interwoven fascicles of spindle cells exhibiting abundant eosinophilic cytoplasm of oncocytic or granular appearance . Mitoses were rarely observed and necrosis was absent . In one case , the advanced lymphocytic infliltration was observed within neoplastic tissue . The tumour cells exhibited immunoreactivity for S-100 protein , galectin-3 , vimentin and epithelial membrane antigen but they were negative for GFAP , anterior pituitary neuroendocrine markers ( prolactin , growth hormone , TSH , ACTH , FSH , LH ) , chromogranin , synaptophysin , cytokeratin CK ( AE1/AE3 ) , smooth muscle actin , desmin , CD34 and CD68 . MIB1 labeling index did not exceed 10% . Ultrastructurally , the tumour cells were rich in mitochondria with lamellar cristae . Moreover , in Case 2 some tumour cells showed a number of giant mitochondria with severely destructed internal matrix . Spindle cell oncocytoma of the anterior pituitary is often misdiagnosed entity of uncertain histogenesis . It should be considered in the differential diagnosis of various sellar-region lesions of oncocytic morphology . OUTPUT: resisting cell death INPUT: In order to determine the in vivo immune response in glioblastoma , monoclonal and polyclonal antibodies specific for inflammatory leukocytes and immunoregulatory products were utilized to stain tissue from four surgical specimens . The more activated the inflammatory cells , the more activated the tumors appeared to be . In the tumor with the largest infiltration ( Case 3 ) , inflammatory cells were stained for interferon-gamma , interleukin-2 , interleukin-1 beta , lymphotoxin , tumor necrosis factor-alpha , and transforming growth factor-beta . The tumor cells also expressed interleukin-1 beta , interleukin-6 , transforming growth factor-beta , tumor necrosis factor-alpha , and prostaglandin E. In contrast , in the tumor with the least inflammatory response ( Case 1 ) , the tumor cells did not express any cytokines . Expression of cytokines by glioma cells was modest in the two cases with modest inflammatory responses . Cellular inflammation , primarily consisting of T cells and macrophages with few or no B cells or natural killer cells , was two- to 15-fold greater outside the tumor than within . In contrast to leukocytes outside the tumor , which were activated and expressing class II major histocompatibility antigens , leukocytes within the tumor parenchyma or at the tumor's edge were negative for these antigens . In the four specimens studied here , the tumor cells themselves were also negative for class II major histocompatibility antigens . These findings , although preliminary , suggest that inflammatory cells within gliomas are inactivated and that glioma cells may increase the expression of immunosuppressive cytokines in response to an increased lymphocyte infiltrate . This observation , if corroborated by more extensive studies , may help to explain the failure of immune treatments in glioblastoma multiforme . OUTPUT: avoiding immune destruction;tumor promoting inflammation INPUT: BACKGROUND Death receptors ( DR ) of the TNF family function as anti-tumor immune effector molecules . Tumor cells , however , often exhibit DR-signaling resistance . Previous studies indicate that radiation can modify gene expression within tumor cells and increase tumor cell sensitivity to immune attack . The aim of this study is to investigate the synergistic effect of sub-lethal doses of ionizing radiation in sensitizing colorectal carcinoma cells to death receptor-mediated apoptosis . METHODOLOGY/PRINCIPAL FINDINGS The ability of radiation to modulate the expression of multiple death receptors ( Fas/CD95 , TRAILR1/DR4 , TRAILR2/DR5 , TNF-R1 and LTβR ) was examined in colorectal tumor cells . The functional significance of sub-lethal doses of radiation in enhancing tumor cell susceptibility to DR-induced apoptosis was determined by in vitro functional sensitivity assays . The longevity of these changes and the underlying molecular mechanism of irradiation in sensitizing diverse colorectal carcinoma cells to death receptor-mediated apoptosis were also examined . We found that radiation increased surface expression of Fas , DR4 and DR5 but not LTβR or TNF-R1 in these cells . Increased expression of DRs was observed 2 days post-irradiation and remained elevated 7-days post irradiation . Sub-lethal tumor cell irradiation alone exhibited minimal cell death , but effectively sensitized three of three colorectal carcinoma cells to both TRAIL and Fas-induced apoptosis , but not LTβR-induced death . Furthermore , radiation-enhanced Fas and TRAIL-induced cell death lasted as long as 5-days post-irradiation . Specific analysis of intracellular sensitizers to apoptosis indicated that while radiation did reduce Bcl-X(L) and c-FLIP protein expression , this reduction did not correlate with the radiation-enhanced sensitivity to Fas and/or TRAIL mediated apoptosis among the three cell types . CONCLUSIONS/SIGNIFICANCE Irradiation of tumor cells can overcome Fas and TRAIL resistance that is long lasting . Overall , results of these investigations suggest that non-lethal doses of radiation can be used to make human tumors more amenable to attack by anti-tumor effector molecules and cells . OUTPUT: resisting cell death INPUT: Switching to a glycolytic metabolism is a rapid adaptation of tumor cells to hypoxia . Although this metabolic conversion may primarily represent a rescue pathway to meet the bioenergetic and biosynthetic demands of proliferating tumor cells , it also creates a gradient of lactate that mirrors the gradient of oxygen in tumors . More than a metabolic waste , the lactate anion is known to participate to cancer aggressiveness , in part through activation of the hypoxia-inducible factor-1 ( HIF-1 ) pathway in tumor cells . Whether lactate may also directly favor HIF-1 activation in endothelial cells ( ECs ) thereby offering a new druggable option to block angiogenesis is however an unanswered question . In this study , we therefore focused on the role in ECs of monocarboxylate transporter 1 ( MCT1 ) that we previously identified to be the main facilitator of lactate uptake in cancer cells . We found that blockade of lactate influx into ECs led to inhibition of HIF-1-dependent angiogenesis . Our demonstration is based on the unprecedented characterization of lactate-induced HIF-1 activation in normoxic ECs and the consecutive increase in vascular endothelial growth factor receptor 2 ( VEGFR2 ) and basic fibroblast growth factor ( bFGF ) expression . Furthermore , using a variety of functional assays including endothelial cell migration and tubulogenesis together with in vivo imaging of tumor angiogenesis through intravital microscopy and immunohistochemistry , we documented that MCT1 blockers could act as bona fide HIF-1 inhibitors leading to anti-angiogenic effects . Together with the previous demonstration of MCT1 being a key regulator of lactate exchange between tumor cells , the current study identifies MCT1 inhibition as a therapeutic modality combining antimetabolic and anti-angiogenic activities . OUTPUT: inducing angiogenesis INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: We analyzed several factors which could influence the immunogenicity of colon tumor cells , using a series of clones derived from a single chemically induced rat adenocarcinoma cell line . These clones display variable tumorigenic potential in syngeneic immunocompetent animals , and it has been established that in this model the tumorigenicity of the cells depends on their ability to escape immune surveillance . The results show an absence of relationship between tumorigenicity and expression of MHC-class-I antigens , cell adhesion to rat fibroblasts or fibroblast extracellular matrix . The secretion of latent and active TGF beta I appeared to be quite variable from one clone to the other , but was unrelated to tumorigenicity . Unexpectedly , some regressive clones produced elevated levels of this cytokine , suggesting that in this model , spontaneous secretion of TGF beta I is not sufficient to impair the immune system of the host . In contrast , the more tumorigenic clones were more resistant than less tumorigenic ones to cytotoxicity mediated by NK or LAK cells . They also showed arrest of cell proliferation after reaching confluence , something not observed in the less tumorigenic clones . Finally , the strongest relationship with tumorigenicity was found for expression of blood-group carbohydrate antigens . Increased expression of blood-group-H antigen and , conversely , decreased expression of beta-galactoside precursors of this antigen correlated with increased tumorigenicity . OUTPUT:

avoiding immune destruction

HoC_dynamic_5_shot74

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: We report the formation , detection , quantitation and structural characterization of products resulting from the adduction of deoxynucleosides ( deoxyadenosine , deoxyguanosine , deoxycytidine and 5-methyldeoxycytidine ) to the catechol estrogens ( CE ) of estrone , estradiol-17beta and estradiol-17 alpha . The crude products are obtained in a one-pot synthesis through oxidation of catechols to quinones and subsequent Michael-type reaction with the deoxynucleosides in acidic medium . In all experiments , adducts are detected by electrospray ionization mass spectrometry analysis after HPLC separation ( LC/ESI/MS(n) ) . The two pyrimidines deoxycytidine and 5-methyldeoxycytidine yield only CE adducts to deoxynucleosides , which correspond to stable adducts on DNA . For purines , the results depend on the CE ( 2,3- or 3,4-catechols ) used , the function and configuration on carbon 17 ( ketone for estrone , alcohol for alpha and beta isomers of estradiol ) , and on the purine itself ( deoxyadenosine or deoxyguanosine ) . Both stable adducts and deglycosylated adducts are formed , and therefore formation of stable adducts on DNA as well as the loss of purines from the DNA strands could be possible . MS(2) and MS(3) experiments prove to be relevant for further structural determinations , enabling in some cases the elucidation of the regiochemistry of adduction on the A and B rings of the steroid moiety . OUTPUT: genomic instability and mutation INPUT: Semi-conservative replication of double-stranded DNA in eukaryotic cells is an asymmetric process involving leading and lagging strand synthesis and different DNA polymerases . We report a study to analyze the effect of these asymmetries when the replication machinery encounters alkylation-induced DNA adducts . The model system is an EBV-derived shuttle vector which replicates in synchrony with the host human cells and carries as marker gene the bacterial gpt gene . A preferential distribution of N-methyl-N-nitrosourea ( MNU)-induced mutations in the non transcribed DNA strand of the shuttle vector pF1-EBV was previously reported . The hypermutated strand was the leading strand . To test whether the different fidelity of DNA polymerases synthesizing the leading and the lagging strands might contribute to MNU-induced mutation distribution the mutagenesis study was repeated on the shuttle vector pTF-EBV which contains the gpt gene in the inverted orientation . We show that the base substitution error rates on an alkylated substrate are similar for the replication of the leading and lagging strands . Moreover , we present evidence that the fidelity of replication opposite O6-methylguanine adducts of both the leading and lagging strands is not affected by the 3 ' flanking base . The preferential targeting of mutations after replication of alkylated DNA is mainly driven by the base at the 5 ' side of the G residues . OUTPUT: genomic instability and mutation INPUT: INTRODUCTION Various agents used in breast cancer chemotherapy provoke DNA double-strand breaks ( DSBs ) . DSB repair competence determines the sensitivity of cells to these agents whereby aberrations in the repair machinery leads to apoptosis . Proteins required for this pathway can be detected as nuclear foci at sites of DNA damage when the pathway is intact . Here we investigate whether focus formation of repair proteins can predict chemosensitivity of breast cancer . METHODS Core needle biopsy specimens were obtained from sixty cases of primary breast cancer before and 18-24 hours after the first cycle of neoadjuvant epirubicin plus cyclophosphamide ( EC ) treatment . Nuclear focus formation of DNA damage repair proteins was immunohistochemically analyzed and compared with tumor response to chemotherapy . RESULTS EC treatment induced nuclear foci of gammaH2AX , conjugated ubiquitin , and Rad51 in a substantial amount of cases . In contrast , BRCA1 foci were observed before treatment in the majority of the cases and only decreased after EC in thirteen cases . The presence of BRCA1- , gammaH2AX- , or Rad51-foci before treatment or the presence of Rad51-foci after treatment was inversely correlated with tumor response to chemotherapy . DNA damage response ( DDR ) competence was further evaluated by considering all four repair indicators together . A high DDR score significantly correlated with low tumor response to EC and EC + docetaxel whereas other clinicopathological factors analyzed did not . CONCLUSIONS High performing DDR focus formation resulted in tumor resistance to DNA damage-inducing chemotherapy . Our results suggested an importance of evaluation of DDR competence to predict breast cancer chemosensitivity , and merits further studying into its usefulness in exclusion of non-responder patients . OUTPUT: genomic instability and mutation INPUT: Despite an increasing incidence of human breast cancer , its etiology remains unknown . Since some environmental chemicals are stored in human breast fat and are rodent mammary carcinogens , determining the genotoxic potential of environmental agents in this key target tissue is important . An assay was developed for detecting genotoxic activity , as unscheduled DNA synthesis ( UDS ) , induced by chemicals and UV radiation in early passage cultures of normal human mammary epithelial cells ( HMEC ) derived from 5 different women . In order to measure UDS in culture , reduction in the percentage of cells in S-phase was accomplished either by depriving the cells of epidermal growth factor and bovine pituitary extract or by contact inhibition of growth . Cultures were incubated with test chemicals for 24 h in the presence of [ 3H]-thymidine . UDS was quantitated autoradiographically as net grains per nucleus ( nuclear grains minus cytoplasmic background , population average ) with &gt ; or = 6 net nuclear grains considered in repair for any individual cell . A positive response was observed with UV radiation , benzo(a)-pyrene , aflatoxin B1 , ethylmethanesulfonate , 1,6-dinitropyrene , 2-acetylaminofluorene , and tobacco smoke condensate but not 7,12-dimethylbenz(a)anthracene or 2,3,7,8-tetrachlorodibenzo-p-dioxin . These results demonstrate that HMEC from all 5 women examined have the ability to metabolize a variety of environmental chemicals to DNA-reactive forms . Furthermore , some chemicals known either to cause mammary cancer in rodents or to be contaminants in human breast tissue are genotoxic in HMEC . A positive response in passage 9 cultures was observed only with direct acting agents , suggesting that HMEC may lose their metabolic capabilities in longer-term cultures . The HMEC UDS assay may be used to address the role of environmental agents in human breast cancer by determining whether chemicals are DNA reactive or metabolized to DNA reactive species in this critical target tissue . OUTPUT: genomic instability and mutation INPUT: OBJECTIVE To explore the adduct characteristics of styrene and DNA . METHODS The adduct reactions between styrene , urinary mandalic acid(MA) , phenylglyoxalic acid(PGA) , mercapturic acid of styrene ( UMA ) and DNA were studied by ultraviolet spectral analysis . The SO-DNA adducts by 32P-post labeled method , the chemical structures of SO-DNA adducts by GC-MS and NMR were also studied . RESULTS SO combined with DNA at O6 , N2 positions of dGMP to form six adducts , but styrene , urinary mandalic acid , phenylglyoxalic acid and mercapturic acid of styrene did not react with DNA to form adduct . CONCLUSIONS Styrene formed adduct with DNA through its active center metabolite--SO after entering the body . SO combined with DNA at O6 , N2 positions of dGMP to form adducts . If these DNA adducts are not repaired or are mis-repaired before cell duplication , the gene mutation and chemical damage would happen . No adduct reactions are seen among other metabolites of styrene . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot75

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Hepatocellular carcinoma ( HCC ) , one of the most frequent and deadliest cancers , has been increasing considerably in the United States . In the absence of a proven effective therapy for HCC , novel chemopreventive strategies are urgently needed to lower the current morbidity and mortality of HCC . Recently , we have reported that resveratrol , a compound present in grapes and red wine , significantly prevents diethylnitrosamine ( DENA)-induced liver tumorigenesis in rats , although the mechanism of action is not completely understood . In the present study , we have examined the underlying mechanisms of resveratrol chemoprevention of hepatocarcinogenesis by investigating the effects of resveratrol on oxidative damage and inflammatory markers during DENA-initiated rat liver carcinogenesis . There was a significant increase in hepatic lipid peroxidation and protein oxidation in carcinogen control animals compared with their normal counterparts at the end of the study ( 20 weeks ) . Elevated expressions of inducible nitric oxide synthase and 3-nitrotyrosine were noticed in the livers of the same animals . Dietary resveratrol ( 50-300 mg/kg ) administered throughout the study reversed all the aforementioned markers in a dose-responsive fashion in rats challenged with DENA . Resveratrol also elevated the protein and mRNA expression of hepatic nuclear factor E2-related factor 2 ( Nrf2 ) . Results of the present investigation provide evidence that attenuation of oxidative stress and suppression of inflammatory response mediated by Nrf2 could be implicated , at least in part , in the chemopreventive effects of this dietary agent against chemically induced hepatic tumorigenesis in rats . The outcome of this study may benefit the development of resveratrol in the prevention and intervention of human HCC . OUTPUT: tumor promoting inflammation INPUT: Liver cancer , predominantly hepatocellular carcinoma ( HCC ) , represents a complex and fatal malignancy driven primarily by oxidative stress and inflammation . Due to dismal prognosis and limited therapeutic intervention , chemoprevention has emerged as a viable approach to reduce the morbidity and mortality of HCC . Pomegranate fruit is a rich source of phytochemicals endowed with potent antioxidant and anti-inflammatory properties . We previously reported that pomegranate phytochemicals inhibit diethylnitrosamine ( DENA)-initiated hepatocarcinogenesis in rats though nuclear factor E2-related factor 2 ( Nrf2)-mediated antioxidant mechanisms . Since Nrf2 also acts as a key mediator of the nuclear factor-kappaB ( NF-κB)-regulated inflammatory pathway , our present study investigated the anti-inflammatory mechanisms of a pomegranate emulsion ( PE ) during DENA-induced rat hepatocarcinogenesis . Rats were administered with PE ( 1 or 10 g/kg ) 4 weeks before and 18 weeks following DENA initiation . There was a significant increase in hepatic expressions of inducible nitric oxide synthase , 3-nitrotyrosine , heat shock protein 70 and 90 , cyclooxygenase-2 and NF-κB in DENA-exposed rat livers . PE dose-dependently suppressed all aforementioned elevated inflammatory markers . A conspicuous finding of this study involves lack of cardiotoxicity of PE as assessed by monitoring cardiac function using noninvasive echocardiography . Our results provide substantial evidence that suppression of the inflammatory cascade through modulation of NF-κB signaling pathway may represent a novel mechanism of liver tumor inhibitory effects of PE against experimental hepatocarcinogenesis . Data presented here coupled with those of our earlier study underline the importance of simultaneously targeting two interconnected molecular circuits , namely , Nrf2-mediated redox signaling and NF-κB-regulated inflammatory pathway , by pomegranate phytoconstituents to achieve chemoprevention of HCC . OUTPUT: tumor promoting inflammation INPUT: Spindle cell oncocytoma ( SCO ) of the pituitary gland is a relatively recently established , very rare subtype of adenohypophysis tumours that was introduced as a distinct clinicopathological entity in the fourth edition of WHO classification of the central nervous system tumours ( 2007 ) . It is non-endocrine neoplasm of the anterior pituitary that occurs in adults and usually follows a benign clinical course , corresponding to WHO grade I. Up to now , pituitary SCO have been reported occasionally and only 14 cases of SCO have been documented in the literature . Because of their rarity , the pathogenesis and natural history of these tumours have not been fully characterized . We report two additional cases of SCO occurring in females aged 63 years ( Case 1 ) and 65 years ( Case 2 ) , who presented with pan-hypopituitarism , headache and visual field defect . In both cases , the magnetic resonance imaging showed solid sellar mass of moderate size with suprasellar extension . The clinical and radiological features suggested non-functioning pituitary macroadenomas without evidence of invasive growth . One patient presented with tumour recurrence 3 years after undergoing the previous surgical removal of tumour , which was initially misdiagnosed as schwannoma . The first tumour was removed by transsphenoidal surgery and the second one by frontal craniotomy . Histologically and immunohistochemically , both tumours displayed the features typical for SCO of the pituitary . They were composed of interwoven fascicles of spindle cells exhibiting abundant eosinophilic cytoplasm of oncocytic or granular appearance . Mitoses were rarely observed and necrosis was absent . In one case , the advanced lymphocytic infliltration was observed within neoplastic tissue . The tumour cells exhibited immunoreactivity for S-100 protein , galectin-3 , vimentin and epithelial membrane antigen but they were negative for GFAP , anterior pituitary neuroendocrine markers ( prolactin , growth hormone , TSH , ACTH , FSH , LH ) , chromogranin , synaptophysin , cytokeratin CK ( AE1/AE3 ) , smooth muscle actin , desmin , CD34 and CD68 . MIB1 labeling index did not exceed 10% . Ultrastructurally , the tumour cells were rich in mitochondria with lamellar cristae . Moreover , in Case 2 some tumour cells showed a number of giant mitochondria with severely destructed internal matrix . Spindle cell oncocytoma of the anterior pituitary is often misdiagnosed entity of uncertain histogenesis . It should be considered in the differential diagnosis of various sellar-region lesions of oncocytic morphology . OUTPUT: resisting cell death INPUT: Helicobacter hepaticus-infected Rag2(-/-) mice emulate many aspects of human inflammatory bowel disease , including the development of colitis and colon cancer . To elucidate mechanisms of inflammation-induced carcinogenesis , we undertook a comprehensive analysis of histopathology , molecular damage , and gene expression changes during disease progression in these mice . Infected mice developed severe colitis and hepatitis by 10wk post-infection , progressing into colon carcinoma by 20wk post-infection , with pronounced pathology in the cecum and proximal colon marked by infiltration of neutrophils and macrophages . Transcriptional profiling revealed decreased expression of DNA repair and oxidative stress response genes in colon , but not in liver . Mass spectrometric analysis revealed higher levels of DNA and RNA damage products in liver compared to colon and infection-induced increases in 5-chlorocytosine in DNA and RNA and hypoxanthine in DNA . Paradoxically , infection was associated with decreased levels of DNA etheno adducts . Levels of nucleic acid damage from the same chemical class were strongly correlated in both liver and colon . The results support a model of inflammation-mediated carcinogenesis involving infiltration of phagocytes and generation of reactive species that cause local molecular damage leading to cell dysfunction , mutation , and cell death . There are strong correlations among histopathology , phagocyte infiltration , and damage chemistry that suggest a major role for neutrophils in inflammation-associated cancer progression . Further , paradoxical changes in nucleic acid damage were observed in tissue- and chemistry-specific patterns . The results also reveal features of cell stress response that point to microbial pathophysiology and mechanisms of cell senescence as important mechanistic links to cancer . OUTPUT: tumor promoting inflammation;genomic instability and mutation;resisting cell death INPUT: A 51-year-old previously healthy man , an ex-smoker , was admitted to the authors ' medical department with a 3-month history of dry cough ; intermittent fever ; painless , ulcerated cutaneous lesions over the trunk and limbs ( Figure 1 ) ; and progressive weight loss . He was of Greek descent . His medical history was remarkable for nasal polyps , which were surgically removed 15 years earlier . Initially , he had been treated with antibiotics , without improvement . Several days before admission , chest radiography revealed pulmonary infiltrates in the left lower lobe . On admission , physical examination revealed a well-orientated man in mild distress , with inspiratory rhonchi at the lower part of the left lung and scattered erythematous nodules of variable size , some of which were ulcerated . Laboratory values were notable for leukopenia , 3.3 x 10(9)/L ; total protein , 5.9 g/dL ; globulin , 2.2 g/dL ; serum glutamic oxaloacetic transaminase , 86 IU/L ; serum glutamic pyruvic transaminase , 71 IU/L ; and lactate dehydrogenase , 519 U/L . Computed tomograph ( CT ) of the chest showed multiple alveolar opacities bilaterally ( Figure 2 ) . Fiberoptic bronchoscopy did not reveal any important pathologic findings . Results of bronchial biopsy , cytology of bronchoalveolar lavage , washing , brushing , and sputum following bronchoscopy were negative . CT of the brai and sinonasal area revealed an abnormal low-density mass in the left nasal area . CT findings of the abdomen were negative , as were results of a bone marrow biopsy . There was no evidence of immunosuppression . The differential diagnosis , considering the evidence described , included granulomatous or infectious diseases , angiocentric lymphoproliferative lesions , and lymphomas . Biopsy of a skin lesion showed lymphoproliferative infiltration of the dermis with a follicular and angiocentric growth pattern and regional epidermal necrosis . Immunohistochemical stains showed that the tumor cell were positive for CD56 and CD3 ( cytoplasmic positivity ) and expressed the cytotoxic proteins T-cell intracellular antigen and granzyme B ( Figure 3 ) They lacked TdT , CD34 , CD7 , CD8 , TCL-1 , and CD123 . Findings from an in situ hybridization study for Epstein-Barr virus were negative . Give this result , molecular analysis ofT-cell receptor ( TCR ) gene rearrangements was performed using polymerase chain reaction-based TCR-gamma gene , wit negative results . The morphology and the immunophenotype were consistent with natural killer/T-cell lymphoma , nasal-type . Nasal involvement must be first excluded to proceed to the diagnosis of nasal-type natural killer-cell lymphoma . Indeed , histologic examination of the nasal mass revealed its polypoid nature . Thus , the authors were led to the diagnosis of extranodal extranasal natural killer/T-cell lymphoma , nasal-type , CD56-positive , Ep stein-Barr virus-negative , TCR-negative . The patient received combination chemotherapy and completed 4 cycles of cyclophosphamide , doxorubicin vincristine , and prednisone every 14 days for 2 months . Skin lesions improved , and there was no fever soon after the initiation of therapy . Reevaluatio after the fourth cycle , however , disclosed pulmonary infiltrations as well as leukemic infiltration of the central nervous system . The patient had receive systemic salvage chemotherapy and intrathecal infusions of methotrexate . Although the lung lesions had diminished at that time , the patient develope paraplegia , his clinical course rapidly deteriorated , and he eventually died . OUTPUT: avoiding immune destruction;resisting cell death INPUT: This is a case report of a 69-year-old woman with sarcomatoid hepatocellular carcinoma ( HCC ) , which was diagnosed clinically as hemangioma . She was first admitted to our university hospital , complaining of general fatigue in December , 1988 , and cholelithiasis and liver cirrhosis with hepatic tumor in Segment 8 were diagnosed . The serum AFP level was within normal range , and the tumor was diagnosed as hemangioma radiologically . She underwent only cholecystectomy and was well without any therapy for the liver tumor up until March in 1991 when she was readmitted to our university hospital due to rapidly progressive liver dysfunction . The size of the liver tumor was unchanged . Despite intensive care , she died of hepatic failure due to cirrhosis in a decompensation state . At autopsy , a well defined yellowish white tumor of 3 cm in maximum diameter was seen in the cirrhotic liver . Although the largest part of the tumor revealed necrosis and hyalinization , a sarcomatoid part composed of spindle-shaped cells was noted in the peripheral portion . In addition , some necrotic ghost cells , probably hepatocellular carcinoma , were also noted . Low molecular cytokeratin , which is always found in HCCs , was seen in spindle-shaped sarcomatoid cells . The liver tumor was diagnosed as sarcomatoid HCC from these pathological findings . We report this histologically unusual HCC with an immunohistochemical study . OUTPUT:

resisting cell death

HoC_dynamic_5_shot76

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Spindle cell oncocytoma ( SCO ) of the pituitary gland is a relatively recently established , very rare subtype of adenohypophysis tumours that was introduced as a distinct clinicopathological entity in the fourth edition of WHO classification of the central nervous system tumours ( 2007 ) . It is non-endocrine neoplasm of the anterior pituitary that occurs in adults and usually follows a benign clinical course , corresponding to WHO grade I. Up to now , pituitary SCO have been reported occasionally and only 14 cases of SCO have been documented in the literature . Because of their rarity , the pathogenesis and natural history of these tumours have not been fully characterized . We report two additional cases of SCO occurring in females aged 63 years ( Case 1 ) and 65 years ( Case 2 ) , who presented with pan-hypopituitarism , headache and visual field defect . In both cases , the magnetic resonance imaging showed solid sellar mass of moderate size with suprasellar extension . The clinical and radiological features suggested non-functioning pituitary macroadenomas without evidence of invasive growth . One patient presented with tumour recurrence 3 years after undergoing the previous surgical removal of tumour , which was initially misdiagnosed as schwannoma . The first tumour was removed by transsphenoidal surgery and the second one by frontal craniotomy . Histologically and immunohistochemically , both tumours displayed the features typical for SCO of the pituitary . They were composed of interwoven fascicles of spindle cells exhibiting abundant eosinophilic cytoplasm of oncocytic or granular appearance . Mitoses were rarely observed and necrosis was absent . In one case , the advanced lymphocytic infliltration was observed within neoplastic tissue . The tumour cells exhibited immunoreactivity for S-100 protein , galectin-3 , vimentin and epithelial membrane antigen but they were negative for GFAP , anterior pituitary neuroendocrine markers ( prolactin , growth hormone , TSH , ACTH , FSH , LH ) , chromogranin , synaptophysin , cytokeratin CK ( AE1/AE3 ) , smooth muscle actin , desmin , CD34 and CD68 . MIB1 labeling index did not exceed 10% . Ultrastructurally , the tumour cells were rich in mitochondria with lamellar cristae . Moreover , in Case 2 some tumour cells showed a number of giant mitochondria with severely destructed internal matrix . Spindle cell oncocytoma of the anterior pituitary is often misdiagnosed entity of uncertain histogenesis . It should be considered in the differential diagnosis of various sellar-region lesions of oncocytic morphology . OUTPUT: resisting cell death INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: INTRODUCTION A subpopulation of cancer cells , tumor-initiating cells , is believed to be the driving force behind tumorigenesis and resistance to radiation and chemotherapy . The persistence of tumor-initiating cells may depend on altered regulation of DNA damage and checkpoint proteins , as well as a reduced propensity to undergo apoptosis or senescence . METHODS To test this hypothesis , we isolated CD24-/low/CD44+ tumor-initiating cells ( as mammospheres ) from MCF-7 breast cancer cells grown in adherent monolayer culture , and carried out a comprehensive comparison of cell death and DNA damage response pathways prior to and after exposure to ionizing radiation in mammospheres and monolayer MCF-7 cells . Single and double-strand break repair was measured by single-cell gel electrophoresis . The latter was also examined by phosphorylation of histone H2AX and formation of 53BP1 and Rad51 foci . Apoptosis was quantified by flow-cytometric analysis of annexin V-binding and senescence was analyzed on the basis of cellular beta-galactosidase activity . We employed the telomeric repeat amplification protocol to quantify telomerase activity . Expression of key DNA repair and cell cycle regulatory proteins was detected and quantified by western blot analysis . RESULTS Our data demonstrate that in comparison to the bulk population of MCF-7 cells ( predominantly CD24+/CD44+ ) , the MCF-7 mammosphere cells benefit from a multifaceted approach to cellular protection relative to that seen in monolayer cells , including a reduced level of reactive oxygen species , a more active DNA single-strand break repair ( SSBR ) pathway , possibly due to a higher level of expression of the key SSBR protein , human AP endonuclease 1 ( Ape1 ) , and a significantly reduced propensity to undergo senescence as a result of increased telomerase activity and a low level of p21 protein expression . No significant difference was seen in the rates of double-strand break repair ( DSBR ) between the two cell types , but DSBR in mammospheres appears to by-pass the need for H2AX phosphorylation . CONCLUSIONS Enhanced survival of MCF-7 tumor-initiating cells in response to ionizing radiation is primarily dependent on an inherent down-regulation of the senescence pathway . Since MCF-7 cells are representative of cancer cells that do not readily undergo apoptosis , consideration of senescence pathways may play a role in targeting stem cells from such tumors . OUTPUT: enabling replicative immortality;evading growth suppressors;genomic instability and mutation;resisting cell death INPUT: The high glucose consumption of tumor cells even in an oxygen-rich environment , referred to as the Warburg effect , has been noted as a nearly universal biochemical characteristic of cancer cells . Targeting the glycolysis pathway has been explored as an anti-cancer therapeutic strategy to eradicate cancer based on this fundamental biochemical property of cancer cells . Oncoproteins such as Akt and c-Myc regulate cell metabolism . Accumulating studies have uncovered various molecular mechanisms by which oncoproteins affect cellular metabolism , raising a concern as to whether targeting glycolysis will be equally effective in treating cancers arising from different oncogenic activities . Here , we established a dual-regulatable FL5.12 pre-B cell line in which myristoylated Akt is expressed under the control of doxycycline , and c-Myc , fused to the hormone-binding domain of the human estrogen receptor , is activated by 4-hydroxytamoxifen . Using this system , we directly compared the effect of these oncoproteins on cell metabolism in an isogenic background . Activation of either Akt or c-Myc leads to the Warburg effect as indicated by increased cellular glucose uptake , glycolysis , and lactate generation . When cells are treated with glycolysis inhibitors , Akt sensitizes cells to apoptosis , whereas c-Myc does not . In contrast , c-Myc but not Akt sensitizes cells to the inhibition of mitochondrial function . This is correlated with enhanced mitochondrial activities in c-Myc cells . Hence , although both Akt and c-Myc promote aerobic glycolysis , they differentially affect mitochondrial functions and render cells susceptible to the perturbation of cellular metabolic programs . OUTPUT: cellular energetics INPUT: The SR/CR mouse phenotype , first described in 1999 in BALB/c and later bred into C57BL/6 mice , is resistant to cancer formation following high doses of cancer cells administered intraperitoneally . The tumor cell targeting and destruction mechanisms have not been identified . By fluorescence-activated cell sorting analysis , the immune response of SR/CR mice after intraperitoneal injection of cancer cells was investigated and compared with parent strain mice . A massive influx of leukocytes into the peritoneal cavity was found . A large fraction of these leukocytes were polymorphonuclear granulocytes , macrophages and natural killer cells . A relative decrease in influx of B-cells compared with controls was demonstrated . Increased proportions of leukocytes belonging to the innate immune system were also demonstrated in splenocytes of SR/CR mice . Cytospins of peritoneal fluid from SR/CR mice after cancer cell injection showed formations of immune cells morphologically resembling polymorphonuclear granulocytes and macrophages adjoining the cancer cells . The results point to the potential involvement of innate immune cells in cancer immunology . Our data support migration of polymorphonuclear granulocytes , macrophages and NK cells into the peritoneum of the SR/CR mouse in response to intraperitoneal injection of S180 cancer cells . The cell composition of spleens of SR/CR mice reflected the differential regulation of the innate immune cells in peritoneal exudates . Both peritoneal exudates and the spleens of SR/CR mice contained decreased proportions of B-cells compared with BALB/c and C57BL/6 mice . We reproduce important aspects of previous published data and further extend them by showing differentially regulated populations of splenocytes including B-lymphocytes in SR/CR mice compared with parent strain controls . Importantly , this differentially regulated immune response of SR/CR mice could not be found in response to challenge with the lymphoma cell line EL-4 . OUTPUT: avoiding immune destruction INPUT: Cells dissociated from spontaneous and transplanted tumours of C3HJax mammary gland have been cultured on polylysine and gelatin substrates . The isolated cells proliferated to form monolayers with high degree of organoid structure as indicated by formation of alveolar cavities . Differences were observed in the cell attachment , growth pattern , number and size of alveolar cavities , cells which lined the cavity and cell morphology on polylysine and gelatin substrates as compared to conventional cell culture plastic surface . On polylysine more than 90% cells attached rapidly , within 15-45 min after plating , with or without serum and formed confluent monolayers marked by presence of large and small alveolar cavities . Multiple interacting cell types took part in organization of the cavity . Cells lining the cavity constantly proliferated and rearranged to expand it . On gelatin , 60-70% cells attached over a period of 6-24 hr in presence of serum and formed confluent monolayers dominated by small alveolar cavities . Cells forming the cavities were epithelial in nature and cavities once formed did not increase in size . Upon subculture , the cell morphology on these substrates was strikingly different . On polylysine , the predominant cell type had numerous irregular microvilli whereas on gelatin , cells had smoother boundaries with a few stunted cytoplasmic extensions . The cell attachment on conventional surface was low , 40-50% . When seeded at high cell density , formation of alveolar cavities was suppressed and at low cell density , cultures were marked by contact inhibition of cells and failure to attain confluence . These results suggest differential behaviour and interaction of mammary tumour epithelium with the substrates used . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot77

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: In this study , laboratory experiments were carried out in order to come to a better understanding of the fate of polycyclic aromatic hydrocarbons ( PAHs ) in the marine environment and especially on their bioaccumulation , biotransformation and genotoxic effects in fish . Juveniles of turbot ( Scophthalmus maximus ) were exposed to PAHs through different routes via ( 1 ) a mixture of dissolved PAHs , ( 2 ) a PAH-polluted sediment and ( 3 ) an oil fuel elutriate . Fish were exposed 4 days followed by a 6-day depuration period . In each experiment , PAH concentrations in the seawater of the tanks were analysed regularly by gas chromatography coupled with mass spectrometry . Muscle and liver samples were also analysed for parent PAH levels and PAH bioconcentration factors were calculated . Biotransformation was evaluated by measuring the levels of PAH metabolites in fish bile . Genotoxicity was assessed by the alkaline comet assay . Regardless of exposure route , the parent PAH concentrations in the liver and muscle showed a peak level 1 day after the beginning of the exposure , followed by a decrease up to the background level towards the end of the experiment , except for the exposure to dissolved PAHs for which levels were relatively low throughout the study . As a consequence , no bioaccumulation was observed in fish tissues at the end of the experiment . In contrast , regardless of exposure routes , a rapid production of biliary metabolites was observed throughout the whole exposure experiment . This was especially true for 1-hydroxypyrene , the major metabolite of pyrene . After 6 days of recovery in clean water , a significant decrease in the total metabolite concentrations occurred in bile . Fish exposed through either route displayed a significant increase in DNA strand breaks after 4 days of exposure , and significant correlations were observed between the level of biliary PAH metabolites and the level of DNA lesions in fish erythrocytes . Overall results indicate that exposure to either a mixture of dissolved PAHs , a PAH-contaminated sediment or a dispersed oil fuel elutriate leads to biotransformation and increase in DNA damage in fish . The quantification of PAH metabolites in bile and DNA damage in erythrocytes appear to be suitable for environmental monitoring of marine pollution either in the case of accidental oil spills or sediment contamination . OUTPUT: genomic instability and mutation INPUT: Prostate cancer is responsible for major deaths globally after lung cancer . However , etiology of prostate cancer is still unknown . Individual risk and incidence of prostate cancer may result from the interaction of genetic susceptibility with exposure to environmental factors such as infectious agents , tobacco , occupational exposure , dietary carcinogens , and/or hormonal imbalances leading to injury of the prostate and to the development of chronic inflammation . About 30% of all human cancers are caused by tobacco smoking and inhaled pollutants . Inflammation is now regarded as an important hallmark of cancer . The present study has been aimed to explore the pro-inflammatory levels in prostate carcinoma patients by examining the serum levels of novel cytokine interleukin-18 ( IL-18 ) expression in tobacco exposed population . A total of 578 ( n = 284 biopsy proven prostate cancer patients , n = 294 controls with and without tobacco exposed population ) were recruited . Serum IL-18 ( Interleukin-18 ) level was done by ELISA . The IL-18 levels between cancer patients and controls within same mode tobacco exposure as tobacco smoking ( overall ) showed significant difference ( P &lt ; 0.0001 ) and further we compared within stratified group , it significantly differ ( P &lt ; 0.0001 ) in bidi and cigarette smoking than control non users . Furthermore , IL-18 levels in tobacco chewers ( overall ) with gutkha and khaini chewers showed significant difference ( P &lt ; 0.01 ) than controls non users . Moreover , the IL-18 levels between cancer patients and controls with in of combined mode chewers smokers and alcohol ( CSA ) , smokers with alcohol showed significant difference ( P &lt ; 0.01 ) than controls . The IL-18 levels also differed significantly ( P &lt ; 0.05 ) with smokers and chewers in higher stages of III and IV , and showed non significant with in lower stages . Tobacco exposure enhance the inflammation in prostate carcinoma patients in stratified group as it have been represented as a risk factors in various cancers , but this study provide further its role that seems to influence inflammation especially in prostate carcinoma . OUTPUT: tumor promoting inflammation INPUT: The results of experimental studies have indicated the pleiotropic effects of statins in organism , e.g. the influence on cell cycle , apoptosis or angiogenesis . In this study , the effects of simvastatin on selected parameters of apoptosis and proliferation in chemocarcinogen-induced mammary tumorigenesis in female rats were determined . Simvastatin was administered dietary at a dose of 18 mg/kg and highly effective dose of 180 mg/kg the entire experiment ( 18 weeks ) . At autopsy mammary tumors were removed and prepared for immunohistochemical and histomorphological analysis . In treated animals ( simvastatin 180 mg/kg ) , significant decrease by 12% in Bcl-2 protein expression and non-significant decrease by 27% of Ki67 protein expression in tumor cells compared to tumor cells in control animals were observed after semiquantitative evaluation . Morphometrical analysis has shown significant proapototic shift in Bcl-2/Bax ratio in tumor cells . In high grade control carcinoma cells , the expression of Ki67 increased by 37% ( non-significantly ) in comparison with control low grade carcinomas . A histomorphological analysis of malignant tumors has revealed a shift from high grade to low grade carcinomas after simvastatin treatment . The noticeable decrease of mammary tumor frequency and incidence in rats after simvastatin treatment was accompanied with antiapoptotic Blc-2 protein decrease and proapoptotic Bax protein increase in this experiment . OUTPUT: resisting cell death INPUT: Breast cancer is the malignant neoplasia with the highest incidence in women worldwide . Chronic oxidative stress and inflammation have been indicated as major mediators during carcinogenesis and cancer progression . Human studies have not considered the complexity of tumor biology during the stages of cancer advance , limiting their clinical application . The purpose of this study was to characterize systemic oxidative stress and immune response parameters in early ( ED ; TNM I and II ) and advanced disease ( AD ; TNM III and IV ) of patients diagnosed with infiltrative ductal carcinoma breast cancer . Oxidative stress parameters were evaluated by plasmatic lipoperoxidation , carbonyl content , thiobarbituric reactive substances ( TBARS ) , nitric oxide levels ( NO ) , total radical antioxidant parameter ( TRAP ) , superoxide dismutase , and catalase activities and GSH levels . Immune evaluation was determined by TNF-α , IL-1β , IL-12 , and IL-10 levels and leukocytes oxidative burst evaluation by chemiluminescence . Tissue damage analysis included heart ( total CK and CKMB ) , liver ( AST , ALT , GGT ) , and renal ( creatinine , urea , and uric acid ) plasmatic markers . C-reactive protein ( CRP ) and iron metabolism were also evaluated . Analysis of the results verified different oxidative stress statuses occur at distinct cancer stages . ED was characterized by reduction in catalase , 8-isoprostanes , and GSH levels , with enhanced lipid peroxidation and TBARS levels . AD exhibited more pronounced oxidative status , with reduction in catalase activity and TRAP , intense lipid peroxidation and high levels of NO , TBARs , and carbonyl content . ED patients presented a Th2 immune pattern , while AD exhibited Th1 status . CRP levels and ferritin were increased in both stages of disease . Leukocytes burst impairment was observed in both the groups . Plasma iron levels were significantly elevated in AD . The data obtained indicated that oxidative stress enhancement and immune response impairment may be necessary to ensure cancer progression to advanced stages and may result from both host and tumor inflammatory mediators . OUTPUT: tumor promoting inflammation INPUT: Food-derived heterocyclic aromatic amines ( HCAs ) have proved to be carcinogenic in both rodents and nonhuman primates . Two different metabolic pathways are suggested for the metabolic activation of HCA . The hepatic pathway proceeds via a two-step process involving N-hydroxylation by cytochrome P4501A2 and subsequent O-acetylation by N-acetyltransferase-2 . An alternative pathway may be of particular interest in extrahepatic tissues and proceeds via one-electron oxidation catalyzed by prostaglandin H synthase ( PHS ) , rendering free-radical metabolites . In this study , we investigated the metabolic activation of two HCAs , 2-amino-3-methylimidazo[4,5-f]quinoline ( IQ ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , by two different enzyme systems in vitro , generating different primary and secondary reactive metabolites . Rat liver S9 mix and PHS were used as the activating system and represent the hepatic and extrahepatic pathways , respectively . Electron-spin resonance spectroscopy showed that both IQ and PhIP exerted inhibiting effects on PHS-mediated formation of hydroxyl radicals during the conversion of arachidonic acid to prostaglandins . Evidence for the formation of HCA free radicals was presented in an indirect way by the formation of glutathione-derived thiyl radicals , with purified PHS as the activating system . Activation by S9 mix did not result in the formation of detectable radical metabolites , showing that the two metabolic routes primarily led to the formation of different metabolites . In all electron-spin resonance experiments , IQ appeared to be more effective than PhIP . In contrasts , DNA adduct analysis by means of ( 32)P-postlabeling showed similar adduct patterns for S9 and PHS in single-stranded and double-stranded salmon testes DNA after incubation with PhIP , indicating the ultimate formation of a common reactive intermediate . For IQ , activation by PHS led to an additional adduct spot that was not present after S9 activation . Furthermore , activation of IQ resulted in higher adduct levels compared with PhIP for both activation pathways . Overall , adduct levels were higher in single-stranded DNA than double-stranded DNA . Our results showed that the hepatic and extrahepatic pathways resulted in different primary metabolites , while the ultimate formation of a similar reactive intermediate for PhIP , possibly an arylnitrenium ion , suggested that both pathways could play an important role in the onset of carcinogenesis . OUTPUT: genomic instability and mutation INPUT: Prevention of environmentally related cancer will be enhanced by the availability of sensitive early warning systems and by improvements in quantitative assessment of human risks . Accordingly , we have carried out a series of molecular epidemiologic studies aimed at validating a panel of biologic markers , including carcinogen-DNA and -protein adducts , sister chromatid exchange , micronucleus formation , DNA strand breaks , and DNA repair capacity . Results from three such studies illustrate the usefulness of these biomarkers in elucidating low-dose-response relationships , correlations between biomarkers , and the range of variation in biomarkers between individuals exposed to similar concentrations of carcinogens . Low-level workplace or ambient exposures to styrene , ethylene oxide , and polycyclic aromatic hydrocarbons ( PAH ) were associated with significant increases in both molecular dose of carcinogens ( adducts ) and various markers of preclinical effects . Correlations between biomarkers varied by exposure . For example , in the styrene study , sister chromatid exchange frequency was not correlated with any of the markers , in contrast to the studies of ethylene oxide and PAH . Significant molecular effects were observed not only in occupationally exposed people but also in residents of an area in Poland characterized by high levels of air pollution . For example , the mean PAH-DNA level in exposed residents ( winter sample ) was 30.4 adducts per 10(8) nucleotides . This level was significantly higher than that of adducts seen in summer samples from the same area ( 4.2/10(8) , or in winter samples from residents of a rural area ( 11.01/10(8) . Significant seasonal variation in PAH-DNA adduct formation in this group was consistent with recorded fluctuations in air pollution levels . Striking interindividual variation was observed in all three exposed populations . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot78

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Metastasis is the most lethal attribute of human malignancy . High-level expression of survivin is involved in both carcinogenesis and angiogenesis in cancer . Previous studies indicate that a mutation of the threonine residue at position 34 ( Thr34Ala ) of survivin generates a dominant-negative mutant that induces apoptosis , inhibits angiogenesis , and suppresses highly metastatic breast carcinoma in mouse models . We investigated the efficacy of gene therapy with a survivin dominant-negative mutant and possible factors related to lymph node metastasis . The metastasis rate was compared between each group in order to find a survivin-targeted therapy against lymphangiogenesis in its earliest stages . We established lymph node metastasis models and treated animals with H22 tumors with Lip-mSurvivinT34A ( Lip-mS ) , Lip-plasmid ( Lip-P ) , or normal saline ( NS ) . Eight days after the last dose , five randomly chosen mice from each group were sacrificed . We detected the apoptotic index , microvessel density ( MVD ) , lymphatic microvessel density ( LMVD ) , and the expression of VEGF-D with immunohistochemistry . After the remaining animals were sacrificed , we compared the tumor-infiltrated lymph nodes in each group . Administration of mSurvivinT34A plasmid complexed with cationic liposome ( DOTAP/chol ) resulted in the efficacious inhibition of tumor growth and lymph node metastasis within the mouse H22 tumor model . These responses were associated with tumor cell apoptosis , and angiogenesis and lymphangiogenesis inhibition . Our results suggested that Lip-mSurvivinT34A induced apoptosis and inhibited tumor angiogenesis and lymphangiogenesis , thus suppressing tumor growth and lymphatic metastasis . The mSurvivinT34A survivin mutant is a promising strategy of gene therapy to inhibit lymphatic metastasis . OUTPUT: activating invasion and metastasis;inducing angiogenesis;resisting cell death INPUT: OBJECTIVES Lymph node metastasis is among the most important prognostic factors for patients with esophageal squamous cell carcinoma after curative esophagectomy ; however , the extent of lymphadenectomy is still controversial . The objective of the present study was to determine the frequency of lymphatic metastases and to study the pattern of lymph node metastasis in a large study population . METHODS The data from 1361 patients with thoracic esophageal squamous cell carcinoma who underwent curative R0 esophagectomy were retrospectively examined . Logistic regression analysis was used to identify the factors associated with lymph node metastasis . RESULTS Of the 1361 patients , 714 ( 52.5% ) were found to have lymph node metastasis . The frequency of lymph node metastasis increased as the tumor invasion increased . Paratracheal nodes were the most frequent metastasis nodes ( 15.9% ) . The frequency of lymph node metastasis was 9.8% in the neck , 18.0% in the upper mediastinum , 18.9% in the middle mediastinum , 11.8% in the lower mediastinum , and 28.4% in the abdomen . Of these 714 patients , 424 ( 31.2% ) presented with 1 field involvement , 255 ( 18.7% ) with 2 fields , and 35 ( 2.6% ) with 3 fields involvement . Logistic regression analysis revealed tumor length ( P<.001 ) , tumor invasion ( P<.001 ) , tumor differentiation ( P=.003 ) , and lymphovascular invasion ( P<.001 ) were risk factors for lymph node metastasis . Tumor location ( P<.001 ) , tumor invasion ( P=.003 ) , lymphovascular invasion ( P=.004 ) , and paratracheal lymph node involvement ( P=.002 ) were identified as risk factors for cervical lymph node metastasis . CONCLUSIONS Metastases were more frequent in the abdomen than in the neck . Total mediastinal and upper abdominal lymphadenectomy should be carefully conducted . Certain factors , such as tumor location , depth of tumor invasion , lymphovascular invasion , and paratracheal lymph node involvement , might be helpful in determining the need to perform cervical lymphadenectomy in individual patients . OUTPUT: activating invasion and metastasis INPUT: Angiogenesis is increased in hematologic malignancies , including non-Hodgkin lymphoma ( NHL ) . Elevated serum levels of two important angiogenic factors , vascular endothelial growth factor ( VEGF ) and basic fibroblast growth factor ( bFGF ) , are associated with a poor prognosis . Immunohistochemistry was used to evaluate 27 patients with NHL and bone marrow involvement ( 17 with low-grade B-cell NHL , including 7 with higher grade transformation ; 6 with intermediate-grade B-cell NHL ; and 4 with T-cell lymphoma ) . Among the 17 patients with low-grade B-cell NHL , results for 7 were positive for VEGF stain ( 41.2% ) , and results were negative for all other stains for VEGF receptors , bFGF , and bFGF receptors . In the 10 patients with intermediate-grade B-cell NHL and T-cell lymphoma , all VEGF staining was positive ( 100% ) , but bFGF staining was only weakly positive in 2 . Staining results for seven patients who had low-grade B-cell NHL with higher grade transformation showed that VEGF staining was positive in large lymphoid cells of 5 patients and in small lymphoid cells of one patient . Staining for the receptors VEGFR-1 and VEGFR-2 was positive in large lymphoid cells in four and two cases , respectively . Staining for bFGF was positive in two cases of large lymphoid cells . We concluded that VEGF , but not bFGF , was associated with higher tumor grading of NHL and high-grade transformation of low-grade lymphoma . OUTPUT: inducing angiogenesis INPUT: New molecularly targeted therapies are needed for childhood ependymoma . Angiogenesis and the PDGFR pathway could be potential therapeutic targets . This study aimed to screen ependymomas for the expression and clinicopathological correlates of angiogenic factors and potential therapeutic targets including VEGFR , endoglin ( CD105 ) , CD34 , CD31 , c-Kit , PDGFR-α and PDGFR-β . Immunohistochemistry for angiogenesis factors and PDGFR-α and β was performed in 24 archival tumor samples from children and adults treated for ependymoma at our institution . CD31 density , CD105 density and pericyte coverage index ( PCI ) were calculated . These findings were correlated with clinical outcome . VEGFR2 was overexpressed in tumor cells in only one out of 24 cases , but was found overexpressed in the vessels in 6 cases . PDGFR-α and β were found to be over-expressed in the ependymoma tumor cells in seven out of 24 cases ( 29.2 % ) . CD31 density , CD105 density and PCI did not correlate with expression of PDGFRs . Overexpression of PDGFR-α and β in tumor cells and overexpression of PDGFR-α in tumor endothelium had prognostic significance and this was maintained in multivariate analysis for overexpression of PDGFR-α in tumor cells ( 2 year progression free survival was 16.7 ± 15.2 for cases with overexpression of PDGFR-α in the tumor vs. 74.5 ± 15.2 for those with low/no expression , hazard ratio = 5.78 , p = 0.04 ) . A number of angiogenic factors are expressed in ependymoma tumor cells and tumor endothelium . Preliminary evidence suggests that the expression of PDGFRs could have a prognostic significance in ependymoma . This data suggests that PDGFRs should be further evaluated as targets using novel PDGFR inhibitors . OUTPUT: inducing angiogenesis INPUT: OBJECTIVE To study the effects of genistein on the proliferation , apoptosis induction and expression of related gene proteins of human colon cancer cells in vitro and in vivo , and its mechanisms of action . METHODS MTT colorimetric assay was used to detect the effects of genistein on the proliferation of human colon adenocarcinoma SW480 cells . Light and transmission electron microscopy were used to study the histological and ultrastructural changes . Flow cytometry was used to determine the effects of genistein on cell cycle and apoptosis . Flow cytometry and immunohistochemistry were used to determine the effects of genistein on apoptosis induction and expression of related gene proteins of colon cancer cells . RESULTS The MTT colorimetric assay showed that genistein inhibited the proliferation of SW480 cells in a dose-dependent and time-dependent manner , and the highest inhibition rate was 60.2% after 80 microg/ml genistein treatment for 72 h . The light microscopy revealed that many genistein-treated cancer cells were shrunken , disrupted , or showing cytoplasmic vacuolization . The electron microscopic examination showed cell shrinkage , nuclear fragmentation and pronounced chromatin condensation , sometimes formed crescent chromatin condensation attached to the nuclear membrane . The results of flow cytometry showed that : after SW480 cells were treated with 0 , 20 , 40 , 80 microg/ml genistein for 48 h , the FI values of PCNA were 1.49 +/- 0.02 , 1.28 +/- 0.04 , 1.14 +/- 0.03 , and 0.93 +/- 0.08 ; the FI values of VEGF were 1.75 +/- 0.02 , 1.34 +/- 0.06 , 1.32 +/- 0.04 , and 1.23 +/- 0.04 ; the fluorescence index ( FI ) values of p21 were 1.26 +/- 0.05 , 1.36 +/- 0.06 , 1.61 +/- 0.03 , and 1.73 +/- 0.03 , respectively . There were statistically significant differences between the control group and each treatment group ( P &lt ; 0.05 or P &lt ; 0.01 ) . The scores of immunohistochemical staining of PCNA and VEGF proteins were decreased , while p21 increased . There were statistically significant differences between the control group and each treatment group ( P &lt ; 0.05 or P &lt ; 0.01 ) . CONCLUSION Genistein can inhibit the growth of colon cancer cells via apoptosis induction and cell cycle arrest at G(2)/M phase . The anti-tumor mechanisms of genistein may be related with the down-regulation of expression of VEGF and PCNA , and up-regulation of the expression of p21 . OUTPUT: resisting cell death;evading growth suppressors;sustaining proliferative signaling;inducing angiogenesis INPUT: OBJECTIVE To evaluate the difference of angiogenic factors PDGF/dThdPase,VEGF expression and microvessel density ( MVD ) in primary hypopharyngeal tumor and metastasis lymph nodes . METHOD The author studied immunohistochemically a series of 48 primary hypopharyngeal carcinoma patients and metastasis lymph nodes were calculated . RESULT The percentage of VEGF was 25.38% in primary tumor and 21.52% in lymph nodes . No significant difference was found . The percentage of PDGF/dThdPase was 29.59% in primary tumor and and 21.2% in lymph nodes . This showed significent difference . VEGF showed significent difference between live and death group(P &lt ; 0.05 ) and among differentiation group ( P &lt ; 0.05 ) . MVD showed significant difference between live and death group , early and late stage group , and T1-2 and T3-4 group ( P &lt ; 0.05 ) . There were statistically significant correlations between the score of PDGF/ dThdPase , or VEGF and the score of MVD respectively . CONCLUSION The present study suggests that there was a correlation between VEGF or PDGF and MVD . VEGF and MVD were possible to be prognostic discriminators in hypopharyngeal carcinoma . PDGF expression in lymph nodes was significant higher than in primary tumors , and MVD expression in primary tumor was significant higher than in lymph nodes . OUTPUT:

inducing angiogenesis

HoC_dynamic_5_shot79

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Human carcinomas are defined by recurrent chromosomal aneuploidies , which result in a tissue-specific distribution of genomic imbalances . In order to develop models for these genome mutations and to determine their role in tumorigenesis , we generated 45 spontaneously transformed murine cell lines from normal epithelial cells derived from bladder , cervix , colon , kidney , lung , and mammary gland . Phenotypic changes , chromosomal aberrations , centrosome number , and telomerase activity were assayed in control uncultured cells and in three subsequent stages of transformation . Supernumerary centrosomes , binucleate cells , and tetraploidy were observed as early as 48 hr after explantation . In addition , telomerase activity increased throughout progression . Live-cell imaging revealed that failure of cytokinesis , not cell fusion , promoted genome duplication . Spectral karyotyping demonstrated that aneuploidy preceded immortalization , consisting predominantly of whole chromosome losses ( 4 , 9 , 12 , 13 , 16 , and Y ) and gains ( 1 , 10 , 15 , and 19 ) . After transformation , focal amplifications of the oncogenes Myc and Mdm2 were frequently detected . Fifty percent of the transformed lines resulted in tumors on injection into immunocompromised mice . The phenotypic and genomic alterations observed in spontaneously transformed murine epithelial cells recapitulated the aberration pattern observed during human carcinogenesis . The dominant aberration of these cell lines was the presence of specific chromosomal aneuploidies . We propose that our newly derived cancer models will be useful tools to dissect the sequential steps of genome mutations during malignant transformation , and also to identify cancer-specific genes , signaling pathways , and the role of chromosomal instability in this process . OUTPUT: enabling replicative immortality;genomic instability and mutation INPUT: Chronic inflammation is a critical component in breast cancer progression . Pro-inflammatory mediators along with growth/survival factors within the tumor microenvironment potentiate the expression of pro-inflammatory cytokines ( IL-1 , IL-6 , TNF-α ) , chemotactic cytokines and their receptors ( CXCR4 , CXCL12 , CXCL8 ) and angiogenic factors ( VEGF ) that often overcome the effect of anti-inflammatory molecules ( IL-4 , IL-10 ) thus evading the host's antitumor immunity . Detailed knowledge , therefore , of the regulatory mechanisms determining cytokine levels is essential to understand the pathogenesis of breast cancer . HIF-1α and NF-κB transcription factors are important players for the establishment of a pro-inflammatory and potentially oncogenic environment . HIF-1α is the key mediator of the cellular response to oxygen deprivation and induces the expression of genes involved in survival and angiogenesis within solid hypoxic tumors . The expression of these genes is often modulated by the p53 tumor suppressor protein that induces apoptosis or cell cycle arrest in neoplastic cells . Functional crosstalk between HIF-1α and p53 pathways mediated by modulators shared between the two transcription factors such as SRC-1 and SIRT-1 differentially regulate the expression of distinct subsets of their target genes under variable stress conditions . In an attempt to shed light on the complex regulatory mechanisms involved in cancer-related inflammation , we investigated the role of the two common p53 and HIF-1α co-regulators SRC-1 and SIRT-1 , in the expression of the highly potent metastatic chemokine receptor CXCR4 . Both SRC-1 and SIRT-1 overexpression in DSFX-treated MCF-7 cells reduced CXCR4 cellular levels implying that both co-regulators are crucial factors in the determination of the metastatic potential of breast cancer cells . OUTPUT: activating invasion and metastasis INPUT: Cellular senescence forms a barrier that inhibits the acquisition of an immortal phenotype , a critical feature in tumorigenesis . The inactivation of multiple pathways that positively regulate senescence are required for immortalization . To identify these pathways in an unbiased manner , we performed DNA microarray analyses to assess the expression of 20,000 genes in human prostate epithelial cells ( HPECs ) passaged to senescence . These gene expression patterns were then compared with those of HPECs immortalized with the human Papillomavirus 16 E7 oncoprotein . Senescent cells display gene expression patterns that reflect their nonproliferative , differentiated phenotype and express secretory proteases and extracellular matrix components . A comparison of genes transcriptionally up-regulated in senescence to those in which expression is significantly down-regulated in immortalized HPECs identified three genes : the chemokine BRAK , DOC1 , and a member of the insulin-like growth factor axis , IGFBP-3 . Expression of these genes is found to be uniformly lost in human prostate cancer cell lines and xenografts , and previously , their inactivation was documented in tumor samples . Thus , these genes may function in novel pathways that regulate senescence and are inactivated during immortalization . These changes may be critical not only in allowing cells to bypass senescence in vitro but in the progression of prostate cancer in vivo . OUTPUT: enabling replicative immortality INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: "Here , we provide the necessary proof of concept , that it is possible to metabolically create a non-permissive or "" hostile "" stromal microenvironment , which actively prevents tumor engraftment in vivo . We developed a novel genetically engineered fibroblast cell line that completely prevents tumor formation in mice , with a 100% protection rate . No host side effects were apparent.This could represent a viable cellular strategy for preventing and treating a variety of human cancers . More specifically , we examined the autocrine and paracrine effects of the cellular delivery of TNF-alpha on breast cancer tumor growth and cancer metabolism . For this purpose , we recombinantly overexpressed TNF-alpha in human breast cancer cells ( MDA-MB-231 ) or human immortalized fibroblasts ( hTERT-BJ1 ) . Our results directly show that TNF-alpha functions as a potent tumor suppressor . Remarkably , TNF-alpha-expressing breast cancer cells were viable , without any significant increases in their basal apoptotic rate . However , after 4 weeks post-implantation , TNF-alpha-expressing breast cancer cells failed to form any tumors in xenografted mice ( 0 tumors/10 injections ) , ultimately conferring 100% protection against tumorigenesis . Similarly , TNF-alpha-overexpressing fibroblasts were also viable , without any increases in apoptosis . Significantly , complete tumor suppression was obtained by co-injecting TNF-alpha expressing stromal fibroblasts with human breast cancer cells , indicating that paracrine cell-mediated delivery of TNF-alpha can also prevent tumor engraftment and growth ( 0 tumors/10 injections ) . Mechanistically , TNF-alpha induced autophagy and mitochondrial dysfunction in both epithelial cancer cells and stromal fibroblasts , preventing energy transfer from the tumor microenvironment , likely "" starving "" the cancer cells to death . In addition , via qRT-PCR analysis of MDA-MB-231 cells , we observed that TNF-alpha mediated the up-regulation of gene transcripts associated with inflammation and senescence [ IL-1-beta , IL-6 , IL-8 , MCP-1 , COX-2 , p21(WAF1/CIP1) ] and downregulated known tumor-promoting genes ( collagen VI and MMP2 ) . Recombinant overexpression of TNF-alpha receptor(s) in MDA-MB-231 cells also significantly reduced tumor growth , but was not as effective as the TNF-alpha ligand itself in preventing tumor growth . Thus , we propose that stromal cell-mediated delivery of TNF-alpha to human tumors [ using transfected fibroblasts or mesenchymal stem cells ( hMSCs) ] may be a novel and effective strategy for the prevention and treatment of human cancers ." OUTPUT: resisting cell death;enabling replicative immortality;tumor promoting inflammation INPUT: Cellular senescence is the genetically programmed cessation of cellular proliferation . We have recently mapped a putative senescence gene(s) on the X chromosome of Chinese hamster embryo ( CHE ) cells . In the present study , we have utilized microcell-mediated chromosome transfer ( microcell fusion ) to test whether : ( i ) the human X chromosome exhibits similar genetic potential to induce senescence and ( ii ) the deletion or inactivation of the X-linked senescence gene(s) in CHE cells is associated with nickel-induced immortalization . A normal CHE or human X chromosome was first introduced into mouse-cell hybrids , then transferred by microcell fusion into a nickel-transformed , immortal male CHE cell line ( Ni-2/TGR ) with an X deletion ( Xq1 ) . Microcell fusion of the normal CHE X chromosome into tumorigenic Ni-2/TGR cells yielded senescence of all X recipient clones . The normal human X chromosome induced dominant senescence of tumorigenic Ni-2/TGR cells in only 17% of the resulting microcell hybrids ( 14/81 ) . Karyotypic analyses of 13 non-senescing human X chromosome-derived microcell hybrid clones revealed that none of these clones retained the complete X. A normal CHE X chromosome induced senescence of 75% of hybrids obtained with another immortal and tumorigenic nickel-transformed male CHE cell line ( Ni-6/TGR ) , which exhibited no visible deletion of the X chromosome , while the normal human X chromosome , only induced senescence in 19% of these hybrids . Transfer of the normal CHE or human X chromosome into spontaneously transformed and tumorigenic cell lines , CHO/TGR or V79/TGR , had little or no effect on their growth . These data suggest that both human and CHE cells possess similar X-linked genetic activities that regulate the process of cellular senescence , and that in Chinese hamster cells nickel-induced immortalization but not that of CHO or V79 cells is associated with inactivation of an X-linked senescence gene . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot80

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Epidermal growth factor ( EGF ) receptor is inversely related to expression of estrogen receptor ( ER ) and progesterone receptor in primary breast tumors and is a negative predictor for response to endocrine therapy . To investigate a possible causal role of EGF receptor expression in breast cancer progression to hormone independence , we have created an experimental cell system . Epidermal growth factor receptor complementary DNA was introduced in estrogen-dependent ZR-75-1 breast cancer cells , and the resulting ZR/HERc cells exhibited a mitogenic response to epidermal growth factor , thus bypassing estrogen dependence . This EGF-induced proliferation could not be inhibited by antiestrogens . In addition , we noted changes in cell morphology and keratin expression of EGF-stimulated ZR/HERc cells , suggestive of an altered differentiation state . Furthermore , intolerance of functional ER and EGF receptor signal transduction pathways in ZR/HERc cells was observed during simultaneous activation , which possibly explains the inverse relationship of ER and EGF receptor expression in primary tumors . In contrast to the parental cells , ZR/HERc cells rapidly progressed to a stable ER-negative phenotype when cultured in the presence of the antiestrogen hydroxy-tamoxifen . These results suggest a possible role for EGF receptor in progression of breast cancer to hormone independence . OUTPUT: sustaining proliferative signaling INPUT: Estrogen receptor α ( ERα ) expression in breast cancer is predictive of response to endocrine therapy ; however , resistance is common in ERα-positive tumors that overexpress the growth factor receptor ERBB2 . Even in the absence of estrogen , ERα can be activated by growth factors , including the epidermal growth factor ( EGF ) . EGF induces a transcriptional program distinct from estrogen ; however , the mechanism of the stimulus-specific response is unknown . Here we show that the EGF-induced ERα genomic targets , its cistromes , are distinct from those induced by estrogen in a process dependent on the transcription factor AP-1 . The EGF-induced ERα cistrome specifically regulates genes found overexpressed in ERBB2-positive human breast cancers . This provides a potential molecular explanation for the endocrine therapy resistance seen in ERα-positive breast cancers that overexpress ERBB2 . These results suggest a central role for ERα in hormone-refractory breast tumors dependent on growth factor pathway activation and favors the development of therapeutic strategies completely antagonizing ERα , as opposed to blocking its estrogen responsiveness alone . OUTPUT: sustaining proliferative signaling INPUT: Ovarian cancer-related angiogenesis is a complex process orchestrated by many positive and negative regulators . Many growth factors are involved in the development of the tumor-associated vasculature , and from these , endocrine gland-derived vascular endothelial growth factor ( EG-VEGF ) seems to play a crucial role . EG-VEGF is the first organ-specific angiogenic factor and its effects are restricted to the endothelial cells of the endocrine glands . Although EG-VEGF was detected in both normal and neoplastic ovaries , its clinical significance remains controversial . In the present study , we analyzed 30 patients with epithelial ovarian cancer , and the immunohistochemical expression of EG-VEGF was compared with the conventional clinico-pathological parameters of prognosis . Neoplastic cells of the ovarian carcinoma expressed EG-VEGF in 73.33% of the cases , as a cytoplasmic granular product of reaction . We found a strong correlation between the expression of EG-VEGF at protein level and tumor stage , grade , and microscopic type . The expression of EG-VEGF was found in patients with stage III and IV , but not in stage II . The majority of serous adenocarcinoma , half of the cases with clear cell carcinoma and two cases with endometrioid carcinoma showed definite expression in tumor cells . No positive reaction was found in the cases with mucinous carcinoma . Our results showed that EG-VEGF expression is an indicator not only of the advanced stage , but also of ovarian cancer progression . Based on these data , we concluded that EG-VEGF expression in tumor cells of the epithelial ovarian cancer is a good marker of unfavorable prognosis and could be an attractive therapeutic target in patients with advanced-stage tumors , refractory conventional chemotherapy . OUTPUT: inducing angiogenesis INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: The progression of prostate cancers ( PCs ) to locally invasive , androgen-independent and metastatic disease states is generally associated with treatment resistance and disease relapse . The present study was undertaken to establish the possibility of using a combination of specific oncogenic products , including epidermal growth factor receptor ( EGFR ) , pAkt , nuclear factor-kappaB ( NF-κB ) and macrophage inhibitory cytokine-1 ( MIC-1 ) as biomarkers and therapeutic targets for optimizing the management of patients with localized PC at earlier disease stages . The immunohistochemical and immunofluorescence data have revealed that the expression levels of EGFR , Ser(473)-pAkt , NF-κB p65 and MIC-1 proteins were significantly enhanced in the same subset of 76 cases of prostatic adenocarcinoma specimens during the disease progression and these biomarkers were expressed in a small subpopulation of CD133(+) PC cells and the bulk tumor mass of CD133(-) PC cells . Importantly , all of these biomarkers were also overexpressed in 80-100% of 30 PC metastasis bone tissue specimens . Moreover , the results have indicated that the EGF-EGFR signaling pathway can provide critical functions for the self-renewal of side population ( SP ) cells endowed with stem cell-like features from highly invasive WPE1-NB26 cells . Of therapeutic interest , the targeting of EGFR , pAkt , NF-κB or MIC-1 was also effective at suppressing the basal and EGF-promoted prostasphere formation by SP WPE1-NB26 cells , inducing disintegration of SP cell-derived prostaspheres and decreasing the viability of SP and non-SP WPE1-NB26 cell fractions . Also , the targeting of these oncogenic products induced the caspase-dependent apoptosis in chemoresistant SP WPE1-NB26 cells and enhanced their sensibility to the cytotoxic effects induced by docetaxel . These findings suggest that the combined use of EGFR , pAkt , NF-κB and/or MIC-1 may represent promising strategies for improving the accuracy of current diagnostic and prognostic methods and efficacy of treatments of PC patients in considering the disease heterogeneity , thereby preventing PC progression to metastatic and lethal disease states . OUTPUT: activating invasion and metastasis;sustaining proliferative signaling;resisting cell death INPUT: We assayed the estrogen and progesterone cytosolic receptors by using the enzyme immunoassay method , the epidermal growth factor ( EGF ) cell surface receptors by using 125I-labeled hormone , and the levels of polyamines ( putrescine , spermine , and spermidine ) by using a high-pressure liquid chromatography ( HPLC ) procedure in neoplastic and surrounding normal tissues of patients with colorectal cancer . Our findings show that mean polyamine levels in neoplastic tissue were approximately two-fold greater than the levels in normal colonic mucosa . Estrogen and progesterone receptorial content in normal mucosa were twofold greater than those in neoplastic tissue . No significant differences in EGF receptors were found between colonic cancer tissue and the surrounding normal tissues . The correlations we found between 1 ) estrogen and polyamine levels and 2 ) estrogen and EGF binding suggest the existence of a modulation of the estrogens on colonic mucosa cell proliferation . Furthermore , there was no significant dependency of polyamine and receptor concentrations from the tumor site , the histologic differentiation , or the age and sex of patients . OUTPUT:

sustaining proliferative signaling

HoC_dynamic_5_shot81

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Telomerase is a ribonucleoprotein enzyme that functions to maintain telomeres , the terminal DNA that protects chromosomal integrity , regulating cellular replicative life span . Telomerase is not expressed in most normal human somatic cells but is active in stabilizing telomeres of certain self-renewing cell populations and most malignant cells , making the enzyme an appealing target for anticancer therapy . We describe here a novel cross-species approach to telomerase inhibition . Ectopic expression of the human telomerase catalytic reverse transcriptase component in murine cells inhibited endogenous murine telomerase activity . Using this approach , telomerase inhibition in immortal murine fibroblasts resulted in critical telomere shortening , leading to slowed proliferation , abnormal morphology , altered cell cycle , and telomere dysfunction with cytogenetic instability , followed by apoptotic cell death . Subpopulations of two telomerase-inhibited clones escaped widespread apoptosis , showing proliferative recovery in culture despite persistently inhibited telomerase activity with progressive telomere shortening and dysfunction . This study , by targeting immortal murine cells for telomerase inhibition , demonstrates the importance of telomerase to murine cell immortalization and telomere maintenance . Moreover , the murine model used here should prove useful in further evaluating telomerase inhibition as an anticancer therapy . OUTPUT: enabling replicative immortality;resisting cell death INPUT: Telomerase is a ribonucleoprotein consisting of a catalytic subunit , the telomerase reverse transcriptase , TERT , and an integrally associated RNA , TR , which contains a template for the synthesis of short repetitive G-rich DNA sequences at the ends of telomeres . Telomerase can repetitively reverse transcribe its short RNA template , acting processively to add multiple telomeric repeats onto the same DNA substrate . The contribution of enzyme processivity to telomere length regulation in human cells is not well characterized . In cancer cells , under homeostatic telomere length-maintenance conditions , telomerase acts processively , while under nonequilibrium conditions , telomerase acts distributively on the shortest telomeres . To investigate the role of increased telomerase processivity on telomere length regulation in human cells with limited lifespan that are dependent on human TERT ( hTERT ) for lifespan extension and immortalization , we mutated the leucine at position 866 in the reverse transcriptase C motif of hTERT to a tyrosine ( L866Y ) , which is the amino acid found at a similar position in HIV-1 reverse transcriptase . We report that , similar to the previously reported ' gain of function ' Tetrahymena telomerase mutant ( L813Y ) , the human telomerase variant displays increased processivity. hTERT-L866Y , like wild-type hTERT can immortalize and extend the lifespan of limited lifespan cells . Moreover , hTERT-L866Y expressing cells display heterogenous telomere lengths , telomere elongation , multiple telomeric signals indicative of fragile sites and replicative stress , and an increase in short telomeres , which is accompanied by telomere trimming events . Our results suggest that telomere length and homeostasis in human cells may be regulated by telomerase enzyme processivity . OUTPUT: enabling replicative immortality INPUT: Human carcinomas are defined by recurrent chromosomal aneuploidies , which result in a tissue-specific distribution of genomic imbalances . In order to develop models for these genome mutations and to determine their role in tumorigenesis , we generated 45 spontaneously transformed murine cell lines from normal epithelial cells derived from bladder , cervix , colon , kidney , lung , and mammary gland . Phenotypic changes , chromosomal aberrations , centrosome number , and telomerase activity were assayed in control uncultured cells and in three subsequent stages of transformation . Supernumerary centrosomes , binucleate cells , and tetraploidy were observed as early as 48 hr after explantation . In addition , telomerase activity increased throughout progression . Live-cell imaging revealed that failure of cytokinesis , not cell fusion , promoted genome duplication . Spectral karyotyping demonstrated that aneuploidy preceded immortalization , consisting predominantly of whole chromosome losses ( 4 , 9 , 12 , 13 , 16 , and Y ) and gains ( 1 , 10 , 15 , and 19 ) . After transformation , focal amplifications of the oncogenes Myc and Mdm2 were frequently detected . Fifty percent of the transformed lines resulted in tumors on injection into immunocompromised mice . The phenotypic and genomic alterations observed in spontaneously transformed murine epithelial cells recapitulated the aberration pattern observed during human carcinogenesis . The dominant aberration of these cell lines was the presence of specific chromosomal aneuploidies . We propose that our newly derived cancer models will be useful tools to dissect the sequential steps of genome mutations during malignant transformation , and also to identify cancer-specific genes , signaling pathways , and the role of chromosomal instability in this process . OUTPUT: enabling replicative immortality;genomic instability and mutation INPUT: Direct experimental evidence implicates telomere erosion as a primary cause of cellular senescence . Using a well characterized model system for breast cancer , we define here the molecular and cellular consequences of adriamycin treatment in breast tumor cells . Cells acutely exposed to adriamycin exhibited an increase in p53 activity , a decline in telomerase activity , and a dramatic increase in beta-galactosidase , a marker of senescence . Inactivation of wild-type p53 resulted in a transition of the cellular response to adriamycin treatment from replicative senescence to delayed apoptosis , demonstrating that p53 plays an integral role in the fate of breast tumor cells treated with DNA-damaging agents . Stable introduction of hTERT , the catalytic protein component of telomerase , into MCF-7 cells caused an increase in telomerase activity and telomere length . Treatment of MCF-7-hTERT cells with adriamycin produced an identical senescence response as controls without signs of telomere shortening , indicating that the senescence after treatment is telomere length-independent . However , we found that exposure to adriamycin resulted in an overrepresentation of cytogenetic changes involving telomeres , showing an altered telomere state induced by adriamycin is probably a causal factor leading to the senescence phenotype . To our knowledge , these data are the first to demonstrate that the mechanism of adriamycin-induced senescence is dependent on both functional p53 and telomere dysfunction rather than overall shortening . OUTPUT: enabling replicative immortality;genomic instability and mutation;resisting cell death INPUT: AIM The study was designed to explore the effects of antisense human telomerase RNA ( ahTR ) on the malignant phenotype of gastric carcinoma cell line SGC-7901 , and its potential role in gene therapy for tumors . METHODS An ahTR eukaryotic expression vector , including the sequence of template region of telomere repeats , was constructed by recombinant technology of molecules and then transfected into gastric carcinoma cell line SGC-7901 by liposome DOTAP . Subsequently , the expression of hTR RNA and ahTR RNA by reverse transcription-polymerase chain reaction , telomerase activity by telomeric repeat amplification protocol-ELISA ( TRAP-ELISA ) , telomere length by Southern blotting , cell morphology under light microscope , cellular proliferation capacity by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay , cell-cycle distribution by flow cytometry , efficiency of clone formation in soft agar , and tumorigenecity in nude mice were examined and evaluated in ahTR-transfected cells , control plasmid pCI-neo transfected cells and their parental cells . RESULTS An ahTR eukaryotic expression vector was constructed and successfully transfected into SGC-7901 cells . The telomerase activity in ahTR-transfected SGC-7901 cells decreased from 100% to approximately 25% , and telomere length in the cells shortened to 3.35 from 4.08 Kb at 60 population doublings . Compared with the parental cells and pCI-neo transfected cells , ahTR-transfected cells displayed some morphological changes , such as decreased atypia , and recovery of contact inhibition and density inhibition under light microscope . Furthermore , ahTR-transfected cells displayed decreased invasive capacity in Borden's chamber invasive model , increased G0/G1 phase rate and apoptotic rate . Surprisingly , ahTR-transfected SGC-7901 cells lost their capacity for clone formation in soft agar and tumorigencity in nude mice . CONCLUSION Antisense-hTR transfection can inhibit the growth of SGC-7901 cells and partially reverse the malignant phenotypes . This study provides an exciting approach for cancer therapy by inhibiting telomerase activity using an antisense gene . OUTPUT: evading growth suppressors;activating invasion and metastasis;resisting cell death INPUT: Loss of telomeric DNA during cell proliferation may play a role in ageing and cancer . Since telomeres permit complete replication of eukaryotic chromosomes and protect their ends from recombination , we have measured telomere length , telomerase activity and chromosome rearrangements in human cells before and after transformation with SV40 or Ad5 . In all mortal populations , telomeres shortened by approximately 65 bp/generation during the lifespan of the cultures . When transformed cells reached crisis , the length of the telomeric TTAGGG repeats was only approximately 1.5 kbp and many dicentric chromosomes were observed . In immortal cells , telomere length and frequency of dicentric chromosomes stabilized after crisis . Telomerase activity was not detectable in control or extended lifespan populations but was present in immortal populations . These results suggest that chromosomes with short ( TTAGGG)n tracts are recombinogenic , critically shortened telomeres may be incompatible with cell proliferation and stabilization of telomere length by telomerase may be required for immortalization . OUTPUT:

enabling replicative immortality

HoC_dynamic_5_shot82

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Many cancer cells metabolize glucose preferentially via pyruvate to lactate instead to CO(2) and H(2)O ( oxidative phosphorylation ) even in the presence of oxygen ( Warburg effect ) . Dichloroacetate ( DCA ) is a drug which is able to shift pyruvate metabolism from lactate to acetyl-CoA ( tricarboxylic acid cycle ) by indirect activation of pyruvate dehydrogenase ( PDH ) . This can subsequently lead to an increased flow of oxygen in the respiratory chain , associated with enhanced generation of reactive oxygen species ( ROS ) which may cause apoptosis . In order to investigate if DCA may be suitable for neuroblastoma therapy , it was investigated on three human neuroblastoma cell lines whether DCA can reduce lactate production and enhance oxygen consumption . The data show , that DCA ( in the low millimolar range ) is able to reduce lactate production , but there was only a slight shift to increased oxygen consumption and almost no effect on cell vitality , proliferation and apoptosis of the three cell lines investigated . Therefore , DCA at low millimolar concentrations seems to be only of minor efficacy for neuroblastoma treatment . OUTPUT: cellular energetics;resisting cell death INPUT: Limited options for the treatment of prostate cancer have spurred the search for new therapies . One innovative approach is the use of targeted alpha therapy ( TAT ) to inhibit cancer growth , using an alpha particle emitting radioisotope such as ( 213)Bi . Because of its short range and high linear energy transfer ( LET ) , alpha-particles may be particularly effective in the treatment of cancer , especially in inhibiting the development of metastatic tumors from micro-metastases . Prostate-specific membrane antigen ( PSMA ) is expressed in prostate cancer cells and the neovasculature of a wide variety of malignant neoplasms including lung , colon , breast and others , but not in normal vascular endothelium . The expression is further increased in higher-grade cancers , metastatic disease and hormone-refractory prostate cancer ( PCA ) . J591 is one of several monoclonal antibodies ( mabs ) to the extracellular domain of PSMA . Chelation of J591 mab with ( 213)Bi forms the alpha-radioimmunoconjugate ( AIC ) . The objective of this preclinical study was to design an injectable AIC to treat human prostate tumors growing subcutaneously in mice . The anti-proliferative effects of AIC against prostate cancer were tested in vitro using the MTS assay and in vivo with the nude mice model . Apoptosis was documented using terminal deoxynucleotidyl transferase [ TdT]-mediated deoxyuridinetriphosphate [ dUTP ] nick end-labeling ( TUNEL ) assay , while proliferative index was assessed using the Ki-67 marker . We show that a very high density of PSMA is expressed in an androgen-dependent human PCA cell line ( LNCaP-LN3 ) and in tumor xenografts from nude mice . We also demonstrate that the AIC extensively inhibits the growth of LN3 cells in vitro in a concentration-dependent fashion , causing the cells to undergo apoptosis . Our in vivo studies showed that a local AIC injection of 50 microCi at 2 days post-cell inoculation gave complete inhibition of tumor growth , whereas results for a non-specific AIC were similar to those for untreated mice . Further , after 1 and 3 weeks post-tumor appearance , a single ( 100 microCi/100 microl ) intra-lesional injection of AIC can inhibit the growth of LN3 tumor xenografts ( volume<100 mm(3) ) in nude mice . Tumors treated with AIC decreased in volume from a mean 46+/-14 mm(3) in the first week or 71+/-15 mm(3) in the third week to non-palpable , while in control mice treated with a non-specific AIC using the same dose , tumor volume increased from 42 to 590 mm(3) . There were no observed side effects of the treatment . Because of its in vitro cytotoxicity and these anti-proliferative properties in vivo , the ( 213)Bi-J591 conjugate has considerable potential as a new therapeutic agent for the treatment of prostate cancer . OUTPUT: resisting cell death INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: We have previously demonstrated the antiproliferative effect of two flavonoids-2,2'-dihydroxychalcone ( DHC ) , a novel synthetic flavonoid , and fisetin , a naturally occurring flavonol-in prostate cancer cells . In this study , we further examine the mechanisms of these compounds on survival and proliferation pathways . DHC and fisetin ( 1-50 microM ) caused a dose-dependent reduction in viability , a concomitant increase in apoptosis in PC3 cells at 72 h , and a decrease in clonogenic survival at 24 h treatment . DHC was considerably more potent than fisetin in these cytotoxicity assays . The mechanism of accelerated cellular senescence was not activated by either compound in PC3 or lymph node carcinoma of the prostate ( LNCaP ) cells . Gene expression alterations in PC3 and LNCaP cells treated with 15 muM DHC and 25 microM fisetin for 6 to 24 h were determined by oligonucleotide array . Amongst the most highly represented functional categories of genes altered by both compounds was the cell cycle category . In total , 100 cell cycle genes were altered by DHC and fisetin including 27 genes with key functions in G2/M phase that were downregulated by both compounds . Other functional categories altered included chromosome organization , apoptosis , and stress response . These results demonstrate the multiple mechanisms of antitumor activity of DHC and fisetin in prostate cancer cells in vitro . OUTPUT: enabling replicative immortality;evading growth suppressors;sustaining proliferative signaling INPUT: When the prostate cancer cells become unresponsive to androgen therapy , resistance to chemotherapy becomes imminent , resulting in high mortality . To combat this situation , lycopodine , a pharmacologically important bioactive component derived from Lycopodium clavatum spores , was tested against hormone sensitive ( LnCaP ) and refractory ( PC3 ) prostate cancer cells in vitro . This study aims to check if lycopodine has demonstrable anti-cancer effects and if it has , to find out the possible mechanism of its action . The MTT assay was performed to evaluate the cytotoxic effect . Depolarization of mitochondrial membrane potential , cell cycle , EGF receptor activity and apoptosis were recorded by FACS ; profiles of different anti- and pro-apoptotic genes and their products were studied by semi-quantitative RT-PCR , indirect-ELISA , western blotting . Drug-DNA interaction was determined by CD spectroscopy . Administration of lycopodine down-regulated the expression of 5-lipoxygenase and the 5-oxo-ETE receptor ( OXE receptor1 ) and EGF receptor , and caused up-regulation of cytochrome c with depolarization of mitochondrial inner membrane potential , without palpable change in p53 activity , resulting in apoptosis , cell arrest at G0/G1 stage and ultimately reduced proliferation of cancer cells ; concomitantly , there was externalization of phosphotidyl serine residues . CD spectroscopic analysis revealed intercalating property of lycopodine with DNA molecule , implicating its ability to block cellular DNA synthesis . The overall results suggest that lycopodine is a promising candidate suitable for therapeutic use as an anti-cancer drug . OUTPUT: resisting cell death;sustaining proliferative signaling INPUT: The glycolytic phenotype is a widespread phenomenon in solid cancer forms , including breast cancer . Dichloroacetate ( DCA ) has recently been proposed as a novel and relatively non-toxic anti-cancer agent that can reverse the glycolytic phenotype in cancer cells through the inhibition of pyruvate dehydrogenase kinase . We have examined the effect of DCA against breast cancer cells , including in a highly metastatic in vivo model . The growth of several breast cancer cell lines was found to be inhibited by DCA in vitro . Further examination of 13762 MAT rat mammary adenocarcinoma cells found that reversal of the glycolytic phenotype by DCA correlated with the inhibition of proliferation without any increase in cell death . This was despite a small but significant increase in caspase 3/7 activity , which may sensitize cancer cells to other apoptotic triggers . In vivo , DCA caused a 58% reduction in the number of lung metastases observed macroscopically after injection of 13762 MAT cells into the tail vein of rats ( P = 0.0001 , n &gt ; or = 9 per group ) . These results demonstrate that DCA has anti-proliferative properties in addition to pro-apoptotic properties , and can be effective against highly metastatic disease in vivo , highlighting its potential for clinical use . OUTPUT:

cellular energetics;resisting cell death;activating invasion and metastasis

HoC_dynamic_5_shot83

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Inflammatory response plays an important role not only in the normal physiology but also in the pathology such as cancers . As chronic inflammations are associated with malignancies , it is important to prevent inflammation-mediated neoplastic formation , promotion and/or progression . One possible intervention will be using cancer chemopreventive agents such as curcumin ( CUR ) , a potent anti-inflammatory and anti-oxidative stress compound . Polyunsaturated fatty acids ( PUFA ) such as docosahexaenoic acid ( DHA ) or eicosapentaenoic acid ( EPA ) are potent anti-inflammatory agents by decreasing the production of inflammatory eicosanoids , cytokines , and reactive oxygen species ( ROS ) . The present study aims at examining whether CUR with DHA or EPA would have synergistic anti-inflammatory effects in RAW 264.7 cells . Non-toxic concentrations of single and combination of the compounds were investigated at 6 , 12 and 24h . The nitric oxide ( NO ) suppression effects were most prominent at 24h . All the combinations of CUR and DHA or EPA with lower concentrations of CUR 5 microM and 25 microM of DHA or EPA were found to have synergistic effects in suppressing LPS-stimulated NO and endogenous NO levels . Importantly , very low doses of CUR 2.5 microM and DHA or EPA of 0.78 microM could synergistically suppress the LPS-induced prostaglandin E(2) ( PGE(2) ) . The combinations were also found to suppress iNOS , COX-2 , 5-lipoxygenase ( 5-LOX ) and cPLA(2) but induce HO-1 . Taken together , the present study clearly shows the synergistic anti-inflammatory as well as anti-oxidative stress effects of CUR and PUFA . OUTPUT: tumor promoting inflammation INPUT: Melittin ( 1 ) is a major polypeptide in honey bee venom that has been used traditionally against chronic inflammation and cancer . However , its molecular mechanism has not been determined . In this study , the antitumor effect of 1 was compared with that of NS398 , a cyclooxygenase-2 ( COX-2 ) inhibitor , in vivo and in vitro . Subcutaneous injection of 1 at 0.5 and 5 mg/kg suppressed significantly vascular endothelial growth factor ( VEGF)-A-transfected highly metastatic Lewis lung cancer ( VEGF-A-hm LLC ) tumor growth by 25% and 57% , respectively . Also , 1 inhibited significantly the number of vessels around VEGF-A-hm LLC cells . The results were superior to those obtained in the mice treated with NS398 . Compound 1 dose-dependently inhibited proliferation and tube formation in human umbilical vein endothelial cells ( VEGF-A-HUVECs ) , without affecting cell viability in native HUVECs . In addition , 1 decreased the expression of VEGF receptor-2 ( VEGFR-2 ) , COX-2 , and prostaglandin E(2) ( PGE(2) ) in VEGF-A-transfected HUVECs . These effects were accompanied by a reduction of the phosphorylation of extracellular signal-regulated kinase 1/2 and c-jun N-terminal kinase , whereas it increased the phosphorylation of p38 mitogen-activated protein kinase ( MAPK ) . SB203580 abolished the downregulation of COX-2 and VEGFR-2 and the inhibition of cell proliferation by 1 . The antitumor activity of 1 may be associated with antiangiogenic actions via inhibiting VEGFR-2 and inflammatory mediators involved in the MAPK signaling pathway . OUTPUT: inducing angiogenesis;sustaining proliferative signaling INPUT: Ferric nitrilotriacetate ( Fe-NTA ) is a potent nephrotoxicant and a renal carcinogen that induces its effect by causing oxidative stress . The present study was undertaken to explore protective effect of silymarin , a flavonolignan from milk thistle ( Silybum marianum ) , against Fe-NTA mediated renal oxidative stress , inflammation and tumor promotion response along with elucidation of the implicated mechanism(s) . Administration of Fe-NTA ( 10 mg/kg bd wt , i.p. ) to Swiss albino mice induced marked oxidative stress in kidney , evident from augmentation in renal metallothionein ( MT ) expression , depletion of glutathione content and activities of antioxidant and phase II metabolizing enzymes , and enhancement in production of aldehyde products such as 4-hydroxy-2-nonenal . Fe-NTA also significantly activated nuclear factor kappa B ( NFkappaB ) and upregulated the expression of downstream genes : cyclooxygenase 2 and inducible nitric oxide synthase and enhancing the production of proinflammatory cytokines : tumor necrosis factor alpha ( TNF-alpha ) and interleukin-6 ( IL-6 ) . However , feeding of 0.5% and 1% silymarin diet conferred a significant protection against Fe-NTA induced oxidative stress and inflammation . It further augmented MT expression , restored the antioxidant armory , ameliorated NFkappaB activation and decreased the expression of proinflammatory mediators . Silymarin also suppressed Fe-NTA induced hyperproliferation in kidney , ameliorating renal ornithine decarboxylase activity and DNA synthesis . From these results , it could be concluded that silymarin markedly protects against chemically induced renal cancer and acts plausibly by virtue of its antioxidant , anti-inflammatory and antiproliferative activities . OUTPUT: tumor promoting inflammation INPUT: Colon cancer is the third most common malignant neoplasm in the world and it remains an important cause of death , especially in western countries . The toxic environmental pollutant , 1 , 2-dimethylhydrazine ( DMH ) , is also a colon-specific carcinogen . Tannic acid ( TA ) is reported to be effective against various types of chemically induced toxicity and also carcinogenesis . In the present study , we evaluated the chemopreventive efficacy of TA against DMH induced colon toxicity in a rat model . Efficacy of TA against the colon toxicity was evaluated in terms of biochemical estimation of antioxidant enzyme activities , lipid peroxidation , histopathological changes and expression of early molecular markers of inflammation and tumor promotion . DMH treatment induced oxidative stress enzymes ( p<0.001 ) and an early inflammatory and tumor promotion response in the colons of Wistar rats . TA treatment prevented deteriorative effects induced by DMH through a protective mechanism that involved reduction of oxidative stress as well as COX-2 , i-NOS , PCNA protein expression levels and TNF-α(p<0.001) release . It could be concluded from our results that TA markedly protects against chemically induced colon toxicity and acts plausibly by virtue of its antioxidant , anti-inflammatory and antiproliferative activities . OUTPUT: tumor promoting inflammation INPUT: Hypoxia is widespread in solid tumors as a consequence of poorly structured tumor-derived neovasculature , which is recognized to play a role in the resistance of cancer cells to chemotherapy . Etoposide ( VP-16 ) , a drug commonly used in chemotherapy , leads to enhanced accumulation of cell populations in G2/M phase and increases levels of apoptosis as a topoisomerase II inhibitor . We evaluated the effects of hypoxia on the response of the neuroblastoma cell line CHP126 to VP-16 , in order to delineate the mechanisms responsible for the hypoxia-induced chemoresistance of this clinically conventional anti-cancer agent , with an insight to determining potential indications in neuroblastoma therapy . In this study , physiological hypoxia was shown to attenuate G2/M arrest and apoptosis induced in CHP126 cells by VP-16 . It suppressed drug-related Cdk1 activity with a less elevation of regulator proteins such as cyclin B1 , Cdk7 and reduced caspase activation and PARP cleavage compared to the efficiency observed in normoxic condition , which were significantly relative with hypoxia-driven inhibition of p53 and p-ERK1/2 activation . These results clearly demonstrated that hypoxia had a protective effect against VP-16-induced cytotoxicity , which is likely to provide a further therapeutic knowledge in neuroblastomas . OUTPUT: evading growth suppressors;resisting cell death;sustaining proliferative signaling INPUT: Polyphenols are widely distributed in plants and known for antioxidant and anti-inflammatory properties . Areca nut , rich in polyphenols , is the major component of betel quid and we have previously shown that the extract of areca nut can induce oxidative stress in vitro . In this study , we have further pinpointed that areca nut extract ( ANE ) contains catechin based procyanidins which range from dimers to decamers and polymers ; this was carried out by HPLC and electrospray ionization/mass spectrometry ( ESI/MS ) . To quantify their antioxidant potential , oligomeric and polymeric procyanidins of ANE were separated and evaluated using the Trolox equivalent antioxidant capacity ( TEAC ) assay . The results clearly demonstrated that the antioxidant capacity of the ANE procyanidins increased with the degree of polymerization . The anti-inflammatory potential of ANE was also tested using 12-O-tetradecanoylphorbol-13-acetate ( TPA)-treated human oral cancer SAS cells . ANE inhibited TPA-induced cyclooxygenase-2 ( COX-2 ) protein expression at low doses , which correlated with the inhibition of ERK phosphorylation in the SAS cells . Furthermore , feeding rats with ANE at 1 and 10mg/kg/day for 5days significantly repressed carrageenan-induced inflammatory exudates and PGE(2) formation . In conclusion , ANE , which contains catechins based oligomeric and polymeric procyanidins , regulates COX-2 expression in vitro and possess anti-inflammatory potential in vivo . OUTPUT:

tumor promoting inflammation

HoC_dynamic_5_shot84

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cigarette smoke ( CS ) is a rich source of radicals , predisposing the cell to oxidative stress resulting in inflammation . Chronic inflammation is a recognized risk factor for carcinogenesis . Cyclooxygenase-2 ( COX-2 ) is a mediator of inflammatory pathway and may , therefore , contribute to carcinogenesis . There are several reports that suggest the association between CS and COX-2 associated risk to cancer . In the present study , we examined the role of celecoxib ( a selective COX-2 inhibitor ) in modulating the oxidative stress caused by CS inhalation in mice . CS exposure for a period of 10 weeks caused oxidative stress in the pulmonary and hepatic tissues , as evident from the increase in lipid peroxidation levels ( LPO ) and decrease in reduced glutathione ( GSH ) levels . Celecoxib ( 125 mg/kg body weight for 8 weeks ) administration to CS inhaling mice reduced the oxidative stress by decreasing the LPO levels and enhancing the GSH levels in comparison to the CS-exposed group . CS exposure repressed the enzymatic antioxidant defense system , as evident from the decrease in catalase ( CAT ) and superoxide dismutase ( SOD ) activities . Co-adminstration of celecoxib considerably reversed the changes in the enzymatic antioxidant defense system . Histopathological studies of lungs showed that CS exposure induced alveolar wall destruction and air space enlargement . In co-treated group , the alveolar septa were thicker than normal with apparent infiltration of inflammatory cells . In CS-exposed group , hepatic tissue exhibited vacuolization and macrophage infiltration . Co-treatment with celecoxib restored the normal histoarchitechture in hepatic tissues of CS inhaling mice . Thus , the present study demonstrated that celecoxib adminstration reduced the oxidative stress-mediated risk to carcinogenesis , due to its ability to boost the antioxidant defense system . OUTPUT: tumor promoting inflammation INPUT: Colon cancer is the third most common malignant neoplasm in the world and it remains an important cause of death , especially in western countries . The toxic environmental pollutant , 1 , 2-dimethylhydrazine ( DMH ) , is also a colon-specific carcinogen . Tannic acid ( TA ) is reported to be effective against various types of chemically induced toxicity and also carcinogenesis . In the present study , we evaluated the chemopreventive efficacy of TA against DMH induced colon toxicity in a rat model . Efficacy of TA against the colon toxicity was evaluated in terms of biochemical estimation of antioxidant enzyme activities , lipid peroxidation , histopathological changes and expression of early molecular markers of inflammation and tumor promotion . DMH treatment induced oxidative stress enzymes ( p<0.001 ) and an early inflammatory and tumor promotion response in the colons of Wistar rats . TA treatment prevented deteriorative effects induced by DMH through a protective mechanism that involved reduction of oxidative stress as well as COX-2 , i-NOS , PCNA protein expression levels and TNF-α(p<0.001) release . It could be concluded from our results that TA markedly protects against chemically induced colon toxicity and acts plausibly by virtue of its antioxidant , anti-inflammatory and antiproliferative activities . OUTPUT: tumor promoting inflammation INPUT: Magnetite iron nanoparticles have been widely used as contrast agents and in thermal therapy for cancer . However , their adverse effects on human health have not been fully investigated . In this study , iron oxide nanoparticles were prepared using inorganic iron chloride ( size : 5.3+/-3.6 nm in phosphate buffered saline , surface charge : 23.14 mV ) , and their inflammatory responses were investigated . When mice were treated with iron oxide nanoparticles ( 250 microg/kg , 500 microg/kg , and 1mg/kg ) by a single intratracheal instillation , the level of intracellular reduced glutathione ( GSH ) was decreased in the cells of bronchoalveolar lavage ( BAL ) fluid . The arrest of cell cycles in G1 phase was observed , but S-phase was significantly decreased . The concentrations of pro-inflammatory cytokines ( IL-1 , TNF-alpha , and IL-6 ) were dose-dependently increased at day 1 after instillation in the BAL fluid and in the blood . During the experimental period of 28 days , pro-inflammatory cytokines ( IL-1 , TNF-alpha , and IL-6 ) , Th0 cytokine ( IL-2 ) , Th1 type cytokine ( IL-12 ) , Th2 type cytokines ( IL-4 and IL-5 ) , TGF-beta , and IgE were also elevated . Expressions of many genes related with inflammation or tissue damage such as heat shock protein , matrix metalloproteinase , tissue inhibitors of metalloproteinases , and serum amyloid A were significantly induced . Formation of microgranuloma , which is one of the indicators for chronic inflammatory response , was observed in the alveolar space . In addition , distribution of B cell and CD8+ T cell in blood lymphocytes was increased at day 28 . Based on the result , iron oxide nanoparticles may subchronic induce inflammatory responses via oxidative stress in mice by a single intratracheal instillation . OUTPUT: sustaining proliferative signaling;tumor promoting inflammation INPUT: The soluble hexavalent chromium Cr ( VI ) used in industrial welding is an environmental contaminant widely recognized to act as a carcinogen , mutagen and teratogen towards humans and animals . The carcinogenic potential of metals is a major issue in defining human health risk from exposure . In the present investigation , 93 welders and 60 control subjects with similar mean ages , smoking prevalences and alcohol consumption were enrolled for DNA damage analysis in blood leucocytes by Micronucleus assay ( MN ) and the Comet assay . DNA repair inhibition was also analyzed by assessing XPD gene polymorphism . Welders showed a significant increase in micronucleated cells compared to controls with respect to their smoking habits and alcohol consumption , age and years of exposure ( P<0.05 ) . Results indicated that the welders had a larger mean comet tail length than that of the controls ( P<0.05 ) . The current study suggested that chronic occupational exposure to Cr ( VI ) during welding could lead to increased levels of DNA damage and repair inhibition . OUTPUT: genomic instability and mutation INPUT: We report that all-trans retinoic acid ( ATRA ) in combination with zoledronic acid has strong synergistic cytotoxic and apoptotic effects against human hormone- and drug-refractory prostate cancer cells , PC-3 and DU-145 , in a time- and dose-dependent manner . We further investigated the effect of the combination treatment on the apoptotic process by both oligoarray and protein array analysis in DU-145 cells , in which the drug combination shows much more strong synergistic effects , as compared to PC-3 cells . Moreover , we have also performed real time-PCR array analysis to validate oligoarray results . We demonstrated that the combination of ATRA and zoledronic acid is a strong inducer of apoptotic related cell death in human androgen-and drug refractory prostate cancer cells DU-145 , at either transcriptional or translational levels . While expression of proapoptotic genes such as tumor necrosis factor receptor superfamily ( TNFRSF ) , Bad , Bax , Fas , FADD are induced with the exposure of the combination , expression of antiapoptotic genes or proteins such as members of inhibitor apoptosis family ( IAPs ) , MCL-1 , LTBR , p53 and bcl-2 are reduced . Because this novel combination treatment has fewer side effects than is generally the case with conventional cytotoxic agents , this regimen may be a good option for treatment of elderly prostate cancer patients . OUTPUT: resisting cell death INPUT: Chromated copper arsenate , which is used worldwide as a wood preservative , can adversely affect human health . Accumulating evidence suggests that chromium ( Cr ) and arsenic ( As ) can potentially disrupt the redox balance and cause respiratory diseases and cancer in humans . The present study was designed to determine the combined toxic effects of these metals in the lungs and to clarify the specific molecules that are stimulated by combined exposure to both metals . Male C57BL/6J mice were intratracheally instilled with arsenate [ As(V) ] , hexavalent chromium [ Cr(VI) ] , or a combination of both metals . Mice were sacrificed 2 days after treatment to collect bronchoalveolar lavage fluid and lung tissue samples . Inflammation , cytotoxicity , apoptosis , and oxidative stress markers were measured . Our results indicated that administration of Cr(VI) alone or in combination with As(V) induced neutrophil-dominant inflammation as well as phosphorylation of mitogen-activated protein kinases ; effects of treatment with As(V) alone were comparatively less potent . By analyzing the production of interleukin-6 and activity of lactate dehydrogenase and caspase , we confirmed that co-treatment intensified pulmonary injury and that it was accompanied by oxidative stress , as confirmed by marked increases in the production of reactive oxygen species , reduced glutathione content , and thioredoxin reductase ( TRXRD ) activity . Expressed mRNA levels of heme oxygenase-1 , glutamylcysteine ligase , glutathione peroxidase 2 , thioredoxin ( TRX ) 1 , and TRXRD1 were also enhanced by co-treatment , whereas treatment with As(V) alone reduced the mRNA expression level of TRX2 . Our data suggest that co-treatment with As(V) exacerbated Cr(VI)-induced pulmonary injury and that this effect may be exerted through a disruption in the balance among several antioxidant genes . OUTPUT:

tumor promoting inflammation

HoC_dynamic_5_shot85

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: A common metabolic change in cancer is the acquisition of glycolytic phenotypes . Increased expression of glycolytic enzymes is considered as one contributing factor . The role of mitochondrial defects in acquisition of glycolytic phenotypes has been postulated but remains controversial . Here we show that functional defects in mitochondrial respiration could be induced by oncogenic H-Ras(Q61L) transformation , even though the mitochondrial contents or mass was not reduced in the transformed cells . First , mitochondrial respiration , as measured by mitochondrial oxygen consumption , was suppressed in NIH-3T3 cells transformed with H-Ras(Q61L) . Second , oligomycin or rotenone did not reduce the cellular ATP levels in the H-Ras(Q61L) transformed cells , suggesting a diminished role of mitochondrial respiration in the cellular energy metabolism . Third , inhibition of glycolysis with iodoacetic acid reduced ATP levels at a much faster rate in H-Ras(Q61L) transformed cells than in the vector control cells . The reduction of cellular ATP levels was reversed by exogenously added pyruvate in the vector control cells but not in H-Ras(Q61L) transformed cells . Finally when compared to the HRas(Q61L) transformed cells , the vector control cells had increased resistance toward glucose deprivation . The increased resistance was dependent on mitochondrial oxidative phosphorylation since rotenone or oligomycin abolished the increased survival of the vector control cells under glucose deprivation . The results also suggest an inability of the H-Ras(Q61L) transformed cells to reactivate mitochondrial respiration under glucose deprivation . Taken together , the data suggest that mitochondrial respiration can be impaired during transformation of NIH-3T3 cells by oncogeneic H-Ras(Q61L) . OUTPUT: cellular energetics INPUT: Cancer cells typically display altered glucose metabolism characterized by a preference of aerobic glycolysis , known as the Warburg effect , which facilitates cell proliferation . Hypoxia-inducible factor ( HIF ) and oncoprotein Myc are two prominent transcription factors that drive glycolysis . Previously , we reported that the estrogen-related receptors ( ERRs ) act as cofactors of HIF and enhance HIF-dependent transcription of glycolytic genes under hypoxia . ERRs are orphan nuclear receptors and key regulators of energy metabolism by orchestrating mitochondrial biogenesis , fatty acid oxidation ( FAO ) and oxidative phosphorylation . Here , we show that ERRs also stimulate glycolysis under normoxia . ERRs directly bind to and activate promoters of many genes encoding glycolytic enzymes , and the ERR-binding sites in such promoters are essential for ERR-mediated transcriptional activation . ERRs interact with Myc , and the two factors synergistically activate transcription of glycolytic genes . Furthermore , overexpression of ERRs increases glycolytic gene expression and lactate production . Conversely , depletion of ERRs in cancer cells reduces expression of glycolytic genes and glucose uptake , resulting in decreased aerobic glycolysis and cell growth . Taken together , these results suggest that ERRs are important transcriptional activators of the glycolytic pathway and contribute to the Warburg effect in cancer cells . OUTPUT: cellular energetics;sustaining proliferative signaling INPUT: PURPOSE This study aims to investigate the role of the aberrant expression of Transkelolase-like 1 ( TKTL1 ) in head and neck squamous cell carcinoma ( HNSCC ) tumorigenesis and to characterize TKTL1 contribution to HNSCC tumorigenesis through aerobic glycolysis and HIF1alpha stabilization . EXPERIMENTAL DESIGN TKTL1 promoter hypomethylation and mRNA/protein aberrant expression were studied in human HNSCC tumor samples and normal mucosas . Oncogenic functions of TKTL1 were examined in HNSCC cell line panels and tumor xenograft models with TKTL1 expression construct . The metabolite levels of fructose-6-phosphate , glyceraldehydes-3-phosphate , pyruvate , lactate , and the levels of HIF1alpha protein and its downsteam glycolytic targets were compared between the TKTL1-expressing and vehicle-expressing HNSCC cells . Meanwhile , the effects of HIF1alpha/glycolytic inhibitors were evaluated on the TKTL1 transfectants . RESULTS TKTL1 exhibits high frequency of promoter hypomethylation in HNSCC tumors compared with the normal mucosas , correlating with its overexpression in HNSCC . Overexpression of TKTL1 in HNSCC cells promoted cellular proliferation and enhanced tumor growth in vitro and in vivo . Overexpression of TKTL1 increased the production of fructose-6-phosphate and glyceraldehyde-3-phosphate , in turn elevating the production of pyruvate and lactate , resulting in the normoxic stabilization of the malignancy-promoting transcription factor HIF1alpha and the upregulation of downstream glycolytic enzymes . Notably , the reduction of TKTL1 expression decreased HIF1alpha accumulation and inhibition with HIF1alpha and/or the glycolysis inhibitor could abrogate the growth effects mediated by TKTL1 overexpression . CONCLUSION TKTL1 is a novel candidate oncogene that is epigenetically activated by aberrant hypomethlation and contributes to a malignant phenotype through altered glycolytic metabolism and HIF1alpha accumulation . OUTPUT: cellular energetics INPUT: Glioma tumors are refractory to conventional treatment . Glioblastoma multiforme is the most aggressive type of primary brain tumors in humans . In this study , we introduce oxidative stress-energy depletion ( OSED ) therapy as a new suggested treatment for glioblastoma . OSED utilizes D-amino acid oxidase ( DAO ) , which is a promising therapeutic protein that induces oxidative stress and apoptosis through generating hydrogen peroxide ( H2O2 ) . OSED combines DAO with 3-bromopyruvate ( 3BP ) , a hexokinase II ( HK II ) inhibitor that interferes with Warburg effect , a metabolic alteration of most tumor cells that is characterized by enhanced aerobic glycolysis . Our data revealed that 3BP induced depletion of energetic capabilities of glioma cells. 3BP induced H2O2 production as a novel mechanism of its action . C6 glioma transfected with DAO and treated with D-serine together with 3BP-sensitized glioma cells to 3BP and decreased markedly proliferation , clonogenic power and viability in a three-dimensional tumor model with lesser effect on normal astrocytes . DAO gene therapy using atelocollagen as an in vivo transfection agent proved effective in a glioma tumor model in Sprague-Dawley ( SD ) rats , especially after combination with 3BP . OSED treatment was safe and tolerable in SD rats . OSED therapy may be a promising therapeutic modality for glioma . OUTPUT: cellular energetics INPUT: Aberrant glucose metabolism characterized by high levels of glycolysis , even in the presence of oxygen , is an important hallmark of cancer . This metabolic reprogramming referred to as the Warburg effect is essential to the survival of tumor cells and provides them with substrates required for biomass generation . Molecular mechanisms responsible for this shift in glucose metabolism remain elusive . As described herein , we found that aberrant expression of the proinflammatory protein transglutaminase 2 ( TG2 ) is an important regulator of the Warburg effect in mammary epithelial cells . Mechanistically , TG2 regulated metabolic reprogramming by constitutively activating nuclear factor ( NF)-κB , which binds to the hypoxia-inducible factor ( HIF)-1α promoter and induces its expression even under normoxic conditions . TG2/NF-κB-induced increase in HIF-1α expression was associated with increased glucose uptake , increased lactate production and decreased oxygen consumption by mitochondria . Experimental suppression of TG2 attenuated HIF-1α expression and reversed downstream events in mammary epithelial cells . Moreover , downregulation of p65/RelA or HIF-1α expression in these cells restored normal glucose uptake , lactate production , mitochondrial respiration and glycolytic protein expression . Our results suggest that aberrant expression of TG2 is a master regulator of metabolic reprogramming and facilitates metabolic alterations in epithelial cells even under normoxic conditions . A TG2-induced shift in glucose metabolism helps breast cancer cells to survive under stressful conditions and promotes their metastatic competence . OUTPUT: cellular energetics;tumor promoting inflammation;activating invasion and metastasis INPUT: UNLABELLED ABC transporters like P-glycoprotein ( P-gp/ABCB1 ) are membrane proteins responsible for the transport of toxic compounds out of non-malignant cells and tumor tissue . AIM To investigate the effect of glycolysis and the tissue redox state on P-gp expression in multicellular tumor spheroids derived from prostate adenocarcinoma cells ( DU-145 ) , glioma cells ( Gli36 ) , and the human cervix carcinoma cell line KB-3-1 transfected with a P-gp-EGFP fusion gene that allows monitoring of P-gp expression in living cells . During cell culture of DU-145 , Gli36 , and KB-3-1 tumor spheroids P-gp expression was observed as well as increased lactate and decreased pyruvate levels and expression of glycolytic enzymes . Inhibition of glycolysis for 24 h by either iodoacetate ( IA ) or 2-deoxy-D-glucose ( 2-DDG ) downregulated P-gp expression which was reversed upon coincubation with the radical scavenger ebselen as shown by semi-quantitative immunohistochemisty in DU-145 and Gli36 tumor spheroids , and by EGFP fluorescence in KB-3-1 tumor spheroids . Consequently endogenous ROS generation in DU-145 tumor spheroids was increased in the presence of either IA or 2-DDG , which was abolished upon coincubation with ebselen . Exogenous addition of pyruvate significantly reduced ROS generation , increased P-gp expression as well as efflux of the P-gp substrate doxorubicin . Doxorubicin transport was significantly blunted by 2-DDG and IA , indicating that inhibition of glycolysis reversed the multidrug resistance phenotype . In summary our data demonstrate that P-gp expression in tumor spheroids is closely related to the glycolytic metabolism of tumor cells and can be downregulated by glycolysis inhibitors via mechanisms that involve changes in the cellular redox state . OUTPUT:

cellular energetics

HoC_dynamic_5_shot86

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Tumor endothelial marker ( TEM ) 5 is an adhesion G-protein-coupled receptor upregulated in endothelial cells during tumor and physiologic angiogenesis . So far , the mechanisms leading to upregulation of TEM5 and its function during angiogenesis have not been identified . Here , we report that TEM5 expression in endothelial cells is induced during capillary-like network formation on Matrigel , during capillary morphogenesis in a three-dimensional collagen I matrix , and upon confluence on a two-dimensional matrix . TEM5 expression was not induced by a variety of soluble angiogenic factors , including VEGF and bFGF , in subconfluent endothelial cells . TEM5 upregulation was blocked by toxin B from Clostridium difficile , an inhibitor of the small GTPases Rho , Rac , and Cdc42 . The Rho inhibitor C3 transferase from Clostridium botulinum did not affect TEM5 expression , whereas the Rac inhibitor NSC23766 suppressed TEM5 upregulation . An excess of the soluble TEM5 extracellular domain or an inhibitory monoclonal TEM5 antibody blocked contact inhibition of endothelial cell proliferation resulting in multilayered islands within the endothelial monolayer and increased vessel density during capillary formation . Based on our results we conclude that TEM5 expression during capillary morphogenesis is induced by the small GTPase Rac and mediates contact inhibition of proliferation in endothelial cells . OUTPUT: inducing angiogenesis;evading growth suppressors INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Cells undergoing malignant transformation often exhibit a shift in cellular metabolism from oxidative phosphorylation to glycolysis . This glycolytic shift , called the Warburg effect , provides a mechanistic basis for targeting glycolysis to suppress carcinogenesis through the use of dietary caloric restriction and energy restriction-mimetic agents ( ERMA ) . We recently reported the development of a novel class of ERMAs that exhibits high potency in eliciting starvation-associated cellular responses and epigenetic changes in cancer cells though glucose uptake inhibition . The lead ERMA in this class , OSU-CG5 , decreases the production of ATP and NADH in LNCaP prostate cancer cells . In this study , we examined the effect of OSU-CG5 on the severity of preneoplastic lesions in male transgenic adenocarcinoma of the mouse prostate ( TRAMP ) mice . Daily oral treatment with OSU-CG5 at 100 mg/kg from 6 to 10 weeks of age resulted in a statistically significant decrease in the weight of urogenital tract and microdissected dorsal , lateral , and anterior prostatic lobes relative to vehicle controls . The suppressive effect of OSU-CG5 was evidenced by marked decreases in Ki67 immunostaining and proliferating cell nuclear antigen ( PCNA ) expression in the prostate . OSU-CG5 treatment was not associated with evidence of systemic toxicity . Microarray analysis indicated a central role for Akt , and Western blot analysis showed reduced phosphorylation and/or expression levels of Akt , Src , androgen receptor , and insulin-like growth factor-1 receptor in prostate lobes . These findings support further investigation of OSU-CG5 as a potential chemopreventive agent . OUTPUT: sustaining proliferative signaling INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: We present a new cell line , EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient . The cells show rapid growth in culture with a doubling time of 16 h and high migration activity . Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition . Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma , whereas no metastasis was observed . Cultured EJ cells produced tissue polypeptide antigen ( IPA ) . Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression . Using the DNA sequencing technique , we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed . This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior , and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium . OUTPUT: activating invasion and metastasis;evading growth suppressors;sustaining proliferative signaling;genomic instability and mutation INPUT: Odontogenic tumors originate from the remains of migrating enamel epithelium after the completion of normal tooth genesis . These enamel epithelium remnants exhibit the ability to recapitulate the events that occur during tooth formation . Several lines of evidence suggest that aberrance in the signaling pathways similar to the ones that are used during tooth development , including the WNT pathway , might be the cause of odontogenic tumorigenesis and maintenance . In this study we demonstrated that WNT5A expression was intense in both the epithelial component of ameloblastomas , the most common epithelial odontogenic tumor , and in this tumor's likely precursor cell , the enamel epithelium located at the cervical loop of normal developing human tooth buds . Additionally , when WNT5A was overexpressed in enamel epithelium cells ( LS-8 ) , the clones expressing high levels of WNT5A ( S ) exhibited characteristics of tumorigenic cells , including growth factor independence , loss of anchorage dependence , loss of contact inhibition , and tumor formation in immunocompromised mice . Moreover , overexpression of WNT5A drastically increased LS-8 cell migration and actin reorganization when compared with controls . Suppression of endogenous WNT5A in LS-8 cells ( AS ) greatly impaired their migration and AS cells failed to form significant actin reorganization and membrane protrusion was rarely seen . Taken together , our data indicate that WNT5A signaling is important in modulating tumorigenic behaviors of enamel epithelium cells in ameloblastomas . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot87

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Chronic inflammation is a critical component in breast cancer progression . Pro-inflammatory mediators along with growth/survival factors within the tumor microenvironment potentiate the expression of pro-inflammatory cytokines ( IL-1 , IL-6 , TNF-α ) , chemotactic cytokines and their receptors ( CXCR4 , CXCL12 , CXCL8 ) and angiogenic factors ( VEGF ) that often overcome the effect of anti-inflammatory molecules ( IL-4 , IL-10 ) thus evading the host's antitumor immunity . Detailed knowledge , therefore , of the regulatory mechanisms determining cytokine levels is essential to understand the pathogenesis of breast cancer . HIF-1α and NF-κB transcription factors are important players for the establishment of a pro-inflammatory and potentially oncogenic environment . HIF-1α is the key mediator of the cellular response to oxygen deprivation and induces the expression of genes involved in survival and angiogenesis within solid hypoxic tumors . The expression of these genes is often modulated by the p53 tumor suppressor protein that induces apoptosis or cell cycle arrest in neoplastic cells . Functional crosstalk between HIF-1α and p53 pathways mediated by modulators shared between the two transcription factors such as SRC-1 and SIRT-1 differentially regulate the expression of distinct subsets of their target genes under variable stress conditions . In an attempt to shed light on the complex regulatory mechanisms involved in cancer-related inflammation , we investigated the role of the two common p53 and HIF-1α co-regulators SRC-1 and SIRT-1 , in the expression of the highly potent metastatic chemokine receptor CXCR4 . Both SRC-1 and SIRT-1 overexpression in DSFX-treated MCF-7 cells reduced CXCR4 cellular levels implying that both co-regulators are crucial factors in the determination of the metastatic potential of breast cancer cells . OUTPUT: activating invasion and metastasis INPUT: A common metabolic change in cancer is the acquisition of glycolytic phenotypes . Increased expression of glycolytic enzymes is considered as one contributing factor . The role of mitochondrial defects in acquisition of glycolytic phenotypes has been postulated but remains controversial . Here we show that functional defects in mitochondrial respiration could be induced by oncogenic H-Ras(Q61L) transformation , even though the mitochondrial contents or mass was not reduced in the transformed cells . First , mitochondrial respiration , as measured by mitochondrial oxygen consumption , was suppressed in NIH-3T3 cells transformed with H-Ras(Q61L) . Second , oligomycin or rotenone did not reduce the cellular ATP levels in the H-Ras(Q61L) transformed cells , suggesting a diminished role of mitochondrial respiration in the cellular energy metabolism . Third , inhibition of glycolysis with iodoacetic acid reduced ATP levels at a much faster rate in H-Ras(Q61L) transformed cells than in the vector control cells . The reduction of cellular ATP levels was reversed by exogenously added pyruvate in the vector control cells but not in H-Ras(Q61L) transformed cells . Finally when compared to the HRas(Q61L) transformed cells , the vector control cells had increased resistance toward glucose deprivation . The increased resistance was dependent on mitochondrial oxidative phosphorylation since rotenone or oligomycin abolished the increased survival of the vector control cells under glucose deprivation . The results also suggest an inability of the H-Ras(Q61L) transformed cells to reactivate mitochondrial respiration under glucose deprivation . Taken together , the data suggest that mitochondrial respiration can be impaired during transformation of NIH-3T3 cells by oncogeneic H-Ras(Q61L) . OUTPUT: cellular energetics INPUT: Signal transducers and activators of transcription 3 ( Stat3 ) is activated by cytokines and growth factors in lung cancers and regulates expression of genes implicated in cell growth , survival , and transformation . Previously , we found that mice with a deletion of the G protein-coupled receptor , family C , group 5 , member a ( Gprc5a ) gene develop lung tumors , indicating that Gprc5a is a tumor suppressor . Herein , we show that epithelial cells from Gprc5a knockout mouse lung ( Gprc5a(-/-) cells ) survive better in vitro in medium deprived of exogenous growth factors and form more colonies in semisolid medium than their counterparts from wild-type mice ( Gprc5a(+/+) cells ) . Stat3 tyrosine 705 phosphorylation and expression of several Stat3-regulated antiapoptotic genes were higher in Gprc5a(-/-) than in Gprc5a(+/+) cells . Both cell types secreted leukemia inhibitory factor ( Lif ) ; however , whereas Stat3 activation was persistent in Gprc5a(-/-) cells , it was transient in Gprc5a(+/+) cells . Lung adenocarcinoma cells isolated from Gprc5a(-/-) mice also exhibited autocrine Lif-mediated Stat3 activation . The level of Socs3 , the endogenous Stat3 inhibitory protein , was higher in Gprc5a(+/+) than in Gprc5a(-/-) cells , and expression of the tumor suppressor stabilized Socs3 . Inhibition of Stat3 signaling in Gprc5a(-/-) normal and cancer cells by the Janus-activated kinase 2 inhibitor AG490 or by a dominant negative Stat3(Y705F) increased starvation-induced apoptosis and inhibited colony formation . These results show that persistent Stat3 activation is important for the survival and transformation of Gprc5a(-/-) lung cells and suggest that the tumor suppressive effects of Gprc5a are mediated , at least in part , by inhibition of Stat3 signaling through Socs3 stabilization . OUTPUT: sustaining proliferative signaling INPUT: Disruption of contact inhibition and serum afflux that occur after a tissue injury activate cell cycle , which then stops when confluence is reached again . Although the events involved in cell cycle entry have been widely documented , those managing cell cycle exit have remained so far ill defined . We have identified that the final stage of wound closure is preceded in keratinocytes by a strong accumulation of miR-483-3p , which acts as a mandatory signal triggering cell cycle arrest when confluence is reached . Blocking miR-483-3p accumulation strongly delays cell cycle exit , maintains cells into a proliferative state and retards their differentiation program . Using two models of cell cycle synchronization ( i.e. mechanical injury and serum addition ) , we show that an ectopic upregulation of miR-483-3p blocks cell cycle progression in early G1 phase . This arrest results from a direct targeting of the CDC25A phosphatase by miR-483-3p , which can be impeded using an anti-miRNA against miR-483-3p or a protector that blocks the complex formation between miR-483-3p and the 3'-untranslated region ( UTR ) of CDC25A transcript . We show that the miRNA-induced silencing of CDC25A increases the tyrosine phosphorylation status of CDK4/6 cyclin-dependent kinases which , in turn , abolishes CDK4/6 capacity to associate with D-type cyclins . This prevents CDK4/6 kinases ' activation , impairs downstream events such as cyclin E stimulation and sequesters cells in early G1 . We propose this new regulatory process of cyclin-CDK association as a general mechanism coupling miRNA-mediated CDC25A invalidation to CDK post-transcriptional modifications and cell cycle control . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Sirtuin proteins regulate diverse cellular pathways that influence genomic stability , metabolism and ageing . SIRT7 is a mammalian sirtuin whose biochemical activity , molecular targets and physiological functions have been unclear . Here we show that SIRT7 is an NAD(+)-dependent H3K18Ac ( acetylated lysine 18 of histone H3 ) deacetylase that stabilizes the transformed state of cancer cells . Genome-wide binding studies reveal that SIRT7 binds to promoters of a specific set of gene targets , where it deacetylates H3K18Ac and promotes transcriptional repression . The spectrum of SIRT7 target genes is defined in part by its interaction with the cancer-associated E26 transformed specific ( ETS ) transcription factor ELK4 , and comprises numerous genes with links to tumour suppression . Notably , selective hypoacetylation of H3K18Ac has been linked to oncogenic transformation , and in patients is associated with aggressive tumour phenotypes and poor prognosis . We find that deacetylation of H3K18Ac by SIRT7 is necessary for maintaining essential features of human cancer cells , including anchorage-independent growth and escape from contact inhibition . Moreover , SIRT7 is necessary for a global hypoacetylation of H3K18Ac associated with cellular transformation by the viral oncoprotein E1A . Finally , SIRT7 depletion markedly reduces the tumorigenicity of human cancer cell xenografts in mice . Together , our work establishes SIRT7 as a highly selective H3K18Ac deacetylase and demonstrates a pivotal role for SIRT7 in chromatin regulation , cellular transformation programs and tumour formation in vivo . OUTPUT: evading growth suppressors INPUT: Histone H3 methylation at lysine 4 ( K4 ) is associated with euchromatic regions and is thought to be important for the transcriptional activation of genes during differentiation . In this study , we found that di- and tri-methylation of histone H3 at K4 and acetylation of histones H3 and H4 from the promoter/enhancer to the transcribed region close to the transcription initiation site of the solute carrier family 2 , member 5 ( SLC2A5 ) gene , and its expression , were induced by differentiation of intestine-like Caco-2 cells . These effects were accompanied by contact inhibition of cell growth of these cells . Furthermore , these modifications were induced by co-treatment with a synthetic glucocorticoid hormone dexamethasone and a p44/42 mitogen-activated protein kinase inhibitor PD89059 . Our results suggest that methylation of histone H3 at K4 and acetylation of histones H3 and H4 are involved in SLC2A5 gene induction associated with intestinal differentiation of Caco-2 cells . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot88

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: BACKGROUND The current staging system provides an anatomical classification of lung tumors ; its secondary purpose is to allow the prognostic stratification of patients into homogeneous groups after surgery . In this work , intratumoral perineural invasion , lymphatic and blood vessel invasion together with the necrosis content of the tumor exclusive of the non-small cell cancer staging system were studied . METHODS During a 4-year period , 152 patients operated for non-small cell lung cancer ( NSCLC ) at our hospital were analyzed . Mean age of patients was 55.7 +/- 10.1 years . RESULTS Overall 5-year survival was 42.2 % . Mediastinal lymph node involvement , tumor size , incomplete resection , pneumonectomy , presence of necrosis and perineural invasion were significant prognosticators ( P = 0.03 , 0.04 , 0.0001 , 0.046 , 0.0246 , &lt ; 0.0001 , respectively ) . Multivariate analysis revealed that N status , perineural invasion , and the presence of necrosis were independent prognostic factors ( P = 0.006 , P = 0.001 , P = 0.001 , respectively ) . Patients who had stage I tumor with necrosis and perineural invasion had a lower survival rate than those with stage IIIA tumor without these histopathological features ( P = 0.04 ) . The presence of these histopathological characteristics in stage IIIA patients was a sign of a poorer prognosis ( P = 0.0001 ) . CONCLUSIONS Perineural invasion and the presence of necrosis independently indicated a dismal prognosis and their prognostic power is comparable to those of the TNM classification . These factors could be candidates for better survival stratification and the indicators of the need for adjuvant therapy in early stage lung cancer patients . OUTPUT: activating invasion and metastasis;resisting cell death INPUT: Limited options for the treatment of prostate cancer have spurred the search for new therapies . One innovative approach is the use of targeted alpha therapy ( TAT ) to inhibit cancer growth , using an alpha particle emitting radioisotope such as ( 213)Bi . Because of its short range and high linear energy transfer ( LET ) , alpha-particles may be particularly effective in the treatment of cancer , especially in inhibiting the development of metastatic tumors from micro-metastases . Prostate-specific membrane antigen ( PSMA ) is expressed in prostate cancer cells and the neovasculature of a wide variety of malignant neoplasms including lung , colon , breast and others , but not in normal vascular endothelium . The expression is further increased in higher-grade cancers , metastatic disease and hormone-refractory prostate cancer ( PCA ) . J591 is one of several monoclonal antibodies ( mabs ) to the extracellular domain of PSMA . Chelation of J591 mab with ( 213)Bi forms the alpha-radioimmunoconjugate ( AIC ) . The objective of this preclinical study was to design an injectable AIC to treat human prostate tumors growing subcutaneously in mice . The anti-proliferative effects of AIC against prostate cancer were tested in vitro using the MTS assay and in vivo with the nude mice model . Apoptosis was documented using terminal deoxynucleotidyl transferase [ TdT]-mediated deoxyuridinetriphosphate [ dUTP ] nick end-labeling ( TUNEL ) assay , while proliferative index was assessed using the Ki-67 marker . We show that a very high density of PSMA is expressed in an androgen-dependent human PCA cell line ( LNCaP-LN3 ) and in tumor xenografts from nude mice . We also demonstrate that the AIC extensively inhibits the growth of LN3 cells in vitro in a concentration-dependent fashion , causing the cells to undergo apoptosis . Our in vivo studies showed that a local AIC injection of 50 microCi at 2 days post-cell inoculation gave complete inhibition of tumor growth , whereas results for a non-specific AIC were similar to those for untreated mice . Further , after 1 and 3 weeks post-tumor appearance , a single ( 100 microCi/100 microl ) intra-lesional injection of AIC can inhibit the growth of LN3 tumor xenografts ( volume<100 mm(3) ) in nude mice . Tumors treated with AIC decreased in volume from a mean 46+/-14 mm(3) in the first week or 71+/-15 mm(3) in the third week to non-palpable , while in control mice treated with a non-specific AIC using the same dose , tumor volume increased from 42 to 590 mm(3) . There were no observed side effects of the treatment . Because of its in vitro cytotoxicity and these anti-proliferative properties in vivo , the ( 213)Bi-J591 conjugate has considerable potential as a new therapeutic agent for the treatment of prostate cancer . OUTPUT: resisting cell death INPUT: Breast cancer is the malignant neoplasia with the highest incidence in women worldwide . Chronic oxidative stress and inflammation have been indicated as major mediators during carcinogenesis and cancer progression . Human studies have not considered the complexity of tumor biology during the stages of cancer advance , limiting their clinical application . The purpose of this study was to characterize systemic oxidative stress and immune response parameters in early ( ED ; TNM I and II ) and advanced disease ( AD ; TNM III and IV ) of patients diagnosed with infiltrative ductal carcinoma breast cancer . Oxidative stress parameters were evaluated by plasmatic lipoperoxidation , carbonyl content , thiobarbituric reactive substances ( TBARS ) , nitric oxide levels ( NO ) , total radical antioxidant parameter ( TRAP ) , superoxide dismutase , and catalase activities and GSH levels . Immune evaluation was determined by TNF-α , IL-1β , IL-12 , and IL-10 levels and leukocytes oxidative burst evaluation by chemiluminescence . Tissue damage analysis included heart ( total CK and CKMB ) , liver ( AST , ALT , GGT ) , and renal ( creatinine , urea , and uric acid ) plasmatic markers . C-reactive protein ( CRP ) and iron metabolism were also evaluated . Analysis of the results verified different oxidative stress statuses occur at distinct cancer stages . ED was characterized by reduction in catalase , 8-isoprostanes , and GSH levels , with enhanced lipid peroxidation and TBARS levels . AD exhibited more pronounced oxidative status , with reduction in catalase activity and TRAP , intense lipid peroxidation and high levels of NO , TBARs , and carbonyl content . ED patients presented a Th2 immune pattern , while AD exhibited Th1 status . CRP levels and ferritin were increased in both stages of disease . Leukocytes burst impairment was observed in both the groups . Plasma iron levels were significantly elevated in AD . The data obtained indicated that oxidative stress enhancement and immune response impairment may be necessary to ensure cancer progression to advanced stages and may result from both host and tumor inflammatory mediators . OUTPUT: tumor promoting inflammation INPUT: Colorectal cancer ( CRC ) arises as the consequence of progressive changes from normal epithelial cells through polyp to tumor , and thus is an useful model for studying metabolic shift . In the present study , we studied the metabolomic profiles using high analyte specific gas chromatography/mass spectrometry ( GC/MS ) and liquid chromatography tandem mass spectrometry ( LC/MS/MS ) to attain a systems-level view of the shift in metabolism in cells progressing along the path to CRC . Colonic tissues including tumor , polyps and adjacent matched normal mucosa from 26 patients with sporadic CRC from freshly isolated resections were used for this study . The metabolic profiles were obtained using GC/MS and LC/MS/MS . Our data suggest there was a distinct profile change of a wide range of metabolites from mucosa to tumor tissues . Various amino acids and lipids in the polyps and tumors were elevated , suggesting higher energy needs for increased cellular proliferation . In contrast , significant depletion of glucose and inositol in polyps revealed that glycolysis may be critical in early tumorigenesis . In addition , the accumulation of hypoxanthine and xanthine , and the decrease of uric acid concentration , suggest that the purine biosynthesis pathway could have been substituted by the salvage pathway in CRC . Further , there was a step-wise reduction of deoxycholic acid concentration from mucosa to tumors . It appears that to gain a growth advantage , cancer cells may adopt alternate metabolic pathways in tumorigenesis and this flexibility allows them to adapt and thrive in harsh environment . OUTPUT: cellular energetics INPUT: BACKGROUND Prostate cancer is the second leading cause of cancer mortality among US men . Epidemiological evidence suggests that high vitamin D status protects men from prostate cancer and the active form of vitamin D , 1alpha,25 dihydroxyvitamin D3 ( 1,25(OH)2D ) has anti-cancer effects in cultured prostate cells . Still , the molecular mechanisms and the gene targets for vitamin D-mediated prostate cancer prevention are unknown . RESULTS We examined the effect of 1,25(OH)2D ( +/- 100 nM , 6 , 24 , 48 h ) on the transcript profile of proliferating RWPE1 cells , an immortalized , non-tumorigenic prostate epithelial cell line that is growth arrested by 1,25(OH)2D ( Affymetrix U133 Plus 2.0 , n = 4/treatment per time and dose ) . Our analysis revealed many transcript level changes at a 5% false detection rate : 6 h , 1571 ( 61% up ) , 24 h , 1816 ( 60% up ) , 48 h , 3566 ( 38% up). 288 transcripts were regulated similarly at all time points ( 182 up , 80 down ) and many of the promoters for these transcripts contained putative vitamin D response elements . Functional analysis by pathway or Gene Set Analysis revealed early suppression of WNT , Notch , NF-kB , and IGF1 signaling . Transcripts related to inflammation were suppressed at 6 h ( e.g . IL-1 pathway ) and suppression of proinflammatory pathways continued at later time points ( e.g . IL-17 and IL-6 pathways ) . There was also evidence for induction of anti-angiogenic pathways and induction of transcripts for protection from oxidative stress or maintenance of cell redox homeostasis at 6 h . CONCLUSIONS Our data reveal of large number of potential new , direct vitamin D target genes relevant to prostate cancer prevention . In addition , our data suggests that rather than having a single strong regulatory effect , vitamin D orchestrates a pattern of changes within prostate epithelial cells that limit or slow carcinogenesis . OUTPUT:

tumor promoting inflammation;inducing angiogenesis

HoC_dynamic_5_shot89

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Liver cancer , predominantly hepatocellular carcinoma ( HCC ) , represents a complex and fatal malignancy driven primarily by oxidative stress and inflammation . Due to dismal prognosis and limited therapeutic intervention , chemoprevention has emerged as a viable approach to reduce the morbidity and mortality of HCC . Pomegranate fruit is a rich source of phytochemicals endowed with potent antioxidant and anti-inflammatory properties . We previously reported that pomegranate phytochemicals inhibit diethylnitrosamine ( DENA)-initiated hepatocarcinogenesis in rats though nuclear factor E2-related factor 2 ( Nrf2)-mediated antioxidant mechanisms . Since Nrf2 also acts as a key mediator of the nuclear factor-kappaB ( NF-κB)-regulated inflammatory pathway , our present study investigated the anti-inflammatory mechanisms of a pomegranate emulsion ( PE ) during DENA-induced rat hepatocarcinogenesis . Rats were administered with PE ( 1 or 10 g/kg ) 4 weeks before and 18 weeks following DENA initiation . There was a significant increase in hepatic expressions of inducible nitric oxide synthase , 3-nitrotyrosine , heat shock protein 70 and 90 , cyclooxygenase-2 and NF-κB in DENA-exposed rat livers . PE dose-dependently suppressed all aforementioned elevated inflammatory markers . A conspicuous finding of this study involves lack of cardiotoxicity of PE as assessed by monitoring cardiac function using noninvasive echocardiography . Our results provide substantial evidence that suppression of the inflammatory cascade through modulation of NF-κB signaling pathway may represent a novel mechanism of liver tumor inhibitory effects of PE against experimental hepatocarcinogenesis . Data presented here coupled with those of our earlier study underline the importance of simultaneously targeting two interconnected molecular circuits , namely , Nrf2-mediated redox signaling and NF-κB-regulated inflammatory pathway , by pomegranate phytoconstituents to achieve chemoprevention of HCC . OUTPUT: tumor promoting inflammation INPUT: Hepatocellular carcinoma ( HCC ) , one of the most frequent and deadliest cancers , has been increasing considerably in the United States . In the absence of a proven effective therapy for HCC , novel chemopreventive strategies are urgently needed to lower the current morbidity and mortality of HCC . Recently , we have reported that resveratrol , a compound present in grapes and red wine , significantly prevents diethylnitrosamine ( DENA)-induced liver tumorigenesis in rats , although the mechanism of action is not completely understood . In the present study , we have examined the underlying mechanisms of resveratrol chemoprevention of hepatocarcinogenesis by investigating the effects of resveratrol on oxidative damage and inflammatory markers during DENA-initiated rat liver carcinogenesis . There was a significant increase in hepatic lipid peroxidation and protein oxidation in carcinogen control animals compared with their normal counterparts at the end of the study ( 20 weeks ) . Elevated expressions of inducible nitric oxide synthase and 3-nitrotyrosine were noticed in the livers of the same animals . Dietary resveratrol ( 50-300 mg/kg ) administered throughout the study reversed all the aforementioned markers in a dose-responsive fashion in rats challenged with DENA . Resveratrol also elevated the protein and mRNA expression of hepatic nuclear factor E2-related factor 2 ( Nrf2 ) . Results of the present investigation provide evidence that attenuation of oxidative stress and suppression of inflammatory response mediated by Nrf2 could be implicated , at least in part , in the chemopreventive effects of this dietary agent against chemically induced hepatic tumorigenesis in rats . The outcome of this study may benefit the development of resveratrol in the prevention and intervention of human HCC . OUTPUT: tumor promoting inflammation INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: Curcumin ( CUR ; diferuloylmethane ) , a rhizome extract of Curcuma Longa L. is commonly used as a food coloring and flavoring agent . Although oriental and Ayurvedic medicines have traditionally used CUR in the treatment of diseases , conventional medicine has just begun to recognize its potential therapeutic value . Numerous recent studies have demonstrated the ability of CUR to halt or prevent certain types of cancer , decrease inflammation , and improve cardiovascular health . However , very few studies have examined its ability to protect against drug-induced organ injury . This study explored whether CUR pre-exposure has the potential to prevent acetaminophen ( APAP)-induced : ( i ) hepatotoxicity , ( ii ) genomic injury , ( iii ) oxidative stress in the liver , and ( iv ) apoptotic and necrotic cell deaths in the liver in vivo . Additional goals were to investigate the interplay of pro- and anti-apoptotic genes and their ultimate impact on various forms of cell death . In order to study the CUR-APAP interaction , male B6C3F1 mice were gavaged with CUR ( 17 mg/kg/day , p.o. ) for 12 days followed by a single APAP exposure ( 400 mg/kg , ip ) . Four groups of animals ( control , CUR , APAP , CUR+APAP ) were sacrificed 24 h after APAP exposure . The results indicated that APAP-induced liver injury associated events as serum ALT ( 80-fold ) , lipid peroxidation ( 357% ) and DNA fragmentation ( 469% ) were markedly reduced to 3-fold , 134% and 162% , respectively , in the CUR+APAP group . The APAP-induced increase in expression of pro-apoptotic genes ( Bax , caspase-3 ) decreased while expression of anti-apoptotic genes ( Bcl-XL ) increased in CUR preexposed mouse livers , and these changes were mirrored in the pattern of apoptotic and necrotic cell deaths . Levels of DNA damage sensor P⁵³ and its counterpart Mdm2 were also analyzed during this interaction . Based on the available literature , and these results , it seems likely that CUR may impart global protection in vivo against drug-induced liver injury by opposing several crucial events instrumental to both apoptosis and necrosis . OUTPUT: resisting cell death;tumor promoting inflammation INPUT: Food-derived heterocyclic aromatic amines ( HCAs ) have proved to be carcinogenic in both rodents and nonhuman primates . Two different metabolic pathways are suggested for the metabolic activation of HCA . The hepatic pathway proceeds via a two-step process involving N-hydroxylation by cytochrome P4501A2 and subsequent O-acetylation by N-acetyltransferase-2 . An alternative pathway may be of particular interest in extrahepatic tissues and proceeds via one-electron oxidation catalyzed by prostaglandin H synthase ( PHS ) , rendering free-radical metabolites . In this study , we investigated the metabolic activation of two HCAs , 2-amino-3-methylimidazo[4,5-f]quinoline ( IQ ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , by two different enzyme systems in vitro , generating different primary and secondary reactive metabolites . Rat liver S9 mix and PHS were used as the activating system and represent the hepatic and extrahepatic pathways , respectively . Electron-spin resonance spectroscopy showed that both IQ and PhIP exerted inhibiting effects on PHS-mediated formation of hydroxyl radicals during the conversion of arachidonic acid to prostaglandins . Evidence for the formation of HCA free radicals was presented in an indirect way by the formation of glutathione-derived thiyl radicals , with purified PHS as the activating system . Activation by S9 mix did not result in the formation of detectable radical metabolites , showing that the two metabolic routes primarily led to the formation of different metabolites . In all electron-spin resonance experiments , IQ appeared to be more effective than PhIP . In contrasts , DNA adduct analysis by means of ( 32)P-postlabeling showed similar adduct patterns for S9 and PHS in single-stranded and double-stranded salmon testes DNA after incubation with PhIP , indicating the ultimate formation of a common reactive intermediate . For IQ , activation by PHS led to an additional adduct spot that was not present after S9 activation . Furthermore , activation of IQ resulted in higher adduct levels compared with PhIP for both activation pathways . Overall , adduct levels were higher in single-stranded DNA than double-stranded DNA . Our results showed that the hepatic and extrahepatic pathways resulted in different primary metabolites , while the ultimate formation of a similar reactive intermediate for PhIP , possibly an arylnitrenium ion , suggested that both pathways could play an important role in the onset of carcinogenesis . OUTPUT: genomic instability and mutation INPUT: BACKGROUND AND AIMS Hepatitis C virus ( HCV)-induced chronic inflammation may induce oxidative stress which could compromise the repair of damaged DNA , rendering cells more susceptible to spontaneous or mutagen-induced alterations , the underlying cause of liver cirrhosis and hepatocellular carcinoma . In the current study we examined the induction of reactive oxygen species ( ROS ) resulting from HCV infection and evaluated its effect on the host DNA damage and repair machinery . METHODS HCV infected human hepatoma cells were analyzed to determine ( i ) ROS , ( ii ) 8-oxoG and ( iii ) DNA glycosylases NEIL1 , NEIL2 , OGG1 . Liver biopsies were analyzed for NEIL1 . RESULTS Human hepatoma cells infected with HCV JFH-1 showed 30-60-fold increases in ROS levels compared to uninfected cells . Levels of the oxidatively modified guanosine base 8-oxoguanine ( 8-oxoG ) were significantly increased sixfold in the HCV-infected cells . Because DNA glycosylases are the enzymes that remove oxidized nucleotides , their expression in HCV-infected cells was analyzed . NEIL1 but not OGG1 or NEIL2 gene expression was impaired in HCV-infected cells . In accordance , we found reduced glycosylase ( NEIL1-specific ) activity in HCV-infected cells . The antioxidant N-acetyl cystein ( NAC ) efficiently reversed the NEIL1 repression by inhibiting ROS induction by HCV . NEIL1 expression was also partly restored when virus-infected cells were treated with interferon ( IFN ) . HCV core and to a lesser extent NS3-4a and NS5A induced ROS , and downregulated NEIL1 expression . Liver biopsy specimens showed significant impairment of NEIL1 levels in HCV-infected patients with advanced liver disease compared to patients with no disease . CONCLUSION Collectively , the data indicate that HCV induction of ROS and perturbation of NEIL1 expression may be mechanistically involved in progression of liver disease and suggest that antioxidant and antiviral therapies can reverse these deleterious effects of HCV in part by restoring function of the DNA repair enzyme/s . OUTPUT:

tumor promoting inflammation;genomic instability and mutation

HoC_dynamic_5_shot90

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Ataxia-telangiectasia mutated ( ATM ) is a cellular damage sensor that coordinates the cell cycle with damage-response checkpoints and DNA repair to preserve genomic integrity . However , ATM also has been implicated in metabolic regulation , and ATM deficiency is associated with elevated reactive oxygen species ( ROS ) . ROS has a central role in many physiological and pathophysiological processes including inflammation and chronic diseases such as atherosclerosis and cancer , underscoring the importance of cellular pathways involved in redox homeostasis . We have identified a cytoplasmic function for ATM that participates in the cellular damage response to ROS . We show that in response to elevated ROS , ATM activates the TSC2 tumor suppressor via the LKB1/AMPK metabolic pathway in the cytoplasm to repress mTORC1 and induce autophagy . Importantly , elevated ROS and dysregulation of mTORC1 in ATM-deficient cells is inhibited by rapamycin , which also rescues lymphomagenesis in Atm-deficient mice . Our results identify a cytoplasmic pathway for ROS-induced ATM activation of TSC2 to regulate mTORC1 signaling and autophagy , identifying an integration node for the cellular damage response with key pathways involved in metabolism , protein synthesis , and cell survival . OUTPUT: resisting cell death;tumor promoting inflammation INPUT: BACKGROUND Cellular and clinical sensitivity to ionizing radiation ( IR ) is determined by DNA double-strand breaks ( DSB ) repair . Here , we investigate the molecular mechanism underlying the extreme response of a head and neck tumor case ( SKX ) to standard radiotherapy . METHODS Immunofluorescence ( IF ) was used for the assessment of DSB repair , Western blot and real-time PCR for protein and mRNA expression , respectively . RESULTS SKX cells exhibited a pronounced radiosensitivity associated with numerous residual γ-H2AX foci after IR . This was not associated with lacking canonical repair proteins . SKX cells did not express any ATM protein . Accordingly , immunoblotting revealed no ATM kinase activity toward substrates such as p-SMC1 , p-CHK2 and p-KAP1 . Sequencing of all 66 exons of ATM showed no mutation . ATM mRNA level was moderately reduced , which could be reverted by 5'-Aza-C treatment but without restoring protein levels . Importantly , we demonstrated a post-transcriptional regulation in SKX cells via 6-fold enhanced levels of miR-421 , which targets the 3'-UTR of ATM mRNA . Transfection of SKX cells with either anti-miR-421 inhibitor or a microRNA-insensitive ATM vector recovered ATM expression and abrogated the hyper-radiosensitivity . CONCLUSION This is the first report describing microRNA-mediated down-regulation of ATM leading to clinically manifest tumor radiosensitivity . OUTPUT: genomic instability and mutation INPUT: Several members of the phosphatidylinositol 3-kinase family play key roles in recognising and responding to damage in DNA , induced by a variety of chemicals and other agents . One of these , ATM , the product of the gene mutated in the human genetic disorder ataxia-telangiectasia ( A-T ) , recognises double strand breaks in DNA caused by ionizing radiation and radiomimetic chemicals . In order to study DNA damage recognition and the abnormalities of genome instability and cancer predisposition that occur in A-T patients , we generated a mouse model expressing a mutant form of Atm corresponding to a common human mutation . In this model , a 9 nucleotide in-frame deletion was introduced into the Atm gene and has been designated Atm-Delta SRI . These animals had a longer lifespan than Atm gene disrupted mice ( Atm(-/-) ) and they developed less thymic lymphomas . A characteristic of the lymphomas appearing in Atm-Delta SRI mice was an increased rate of apoptosis compared to the corresponding tumours in Atm(-/-) mice . Increased expression of FasL in these tumours may account for the higher levels of apoptosis . These results demonstrate that expression of mutant Atm in mice gives rise to phenotypic differences compared to Atm(-/-) mice and has implications for heterogeneity described in the human syndrome . OUTPUT: genomic instability and mutation;resisting cell death INPUT: The ATM/ATR DNA damage checkpoint functions in the maintenance of genetic stability and some missense variants of the ATM gene have been shown to confer a moderate increased risk of prostate cancer . However , whether inactivation of this checkpoint contributes directly to prostate specific cancer predisposition is still unknown . Here , we show that exposure of non-malignant prostate epithelial cells ( HPr-1AR ) to androgen led to activation of the ATM/ATR DNA damage response and induction of cellular senescence . Notably , knockdown of the ATM gene expression in HPr-1AR cells can promote androgen-induced TMPRSS2 : ERG rearrangement , a prostate-specific chromosome translocation frequently found in prostate cancer cells . Intriguingly , unlike the non-malignant prostate epithelial cells , the ATM/ATR DNA damage checkpoint appears to be defective in prostate cancer cells , since androgen treatment only induced a partial activation of the DNA damage response . This mechanism appears to preserve androgen induced autophosphorylation of ATM and phosphorylation of H2AX , lesion processing and repair pathway yet restrain ATM/CHK1/CHK2 and p53 signaling pathway . Our findings demonstrate that ATM/ATR inactivation is a crucial step in promoting androgen-induced genomic instability and prostate carcinogenesis . OUTPUT: enabling replicative immortality;genomic instability and mutation INPUT: AIM The study was designed to explore the effects of antisense human telomerase RNA ( ahTR ) on the malignant phenotype of gastric carcinoma cell line SGC-7901 , and its potential role in gene therapy for tumors . METHODS An ahTR eukaryotic expression vector , including the sequence of template region of telomere repeats , was constructed by recombinant technology of molecules and then transfected into gastric carcinoma cell line SGC-7901 by liposome DOTAP . Subsequently , the expression of hTR RNA and ahTR RNA by reverse transcription-polymerase chain reaction , telomerase activity by telomeric repeat amplification protocol-ELISA ( TRAP-ELISA ) , telomere length by Southern blotting , cell morphology under light microscope , cellular proliferation capacity by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay , cell-cycle distribution by flow cytometry , efficiency of clone formation in soft agar , and tumorigenecity in nude mice were examined and evaluated in ahTR-transfected cells , control plasmid pCI-neo transfected cells and their parental cells . RESULTS An ahTR eukaryotic expression vector was constructed and successfully transfected into SGC-7901 cells . The telomerase activity in ahTR-transfected SGC-7901 cells decreased from 100% to approximately 25% , and telomere length in the cells shortened to 3.35 from 4.08 Kb at 60 population doublings . Compared with the parental cells and pCI-neo transfected cells , ahTR-transfected cells displayed some morphological changes , such as decreased atypia , and recovery of contact inhibition and density inhibition under light microscope . Furthermore , ahTR-transfected cells displayed decreased invasive capacity in Borden's chamber invasive model , increased G0/G1 phase rate and apoptotic rate . Surprisingly , ahTR-transfected SGC-7901 cells lost their capacity for clone formation in soft agar and tumorigencity in nude mice . CONCLUSION Antisense-hTR transfection can inhibit the growth of SGC-7901 cells and partially reverse the malignant phenotypes . This study provides an exciting approach for cancer therapy by inhibiting telomerase activity using an antisense gene . OUTPUT: evading growth suppressors;activating invasion and metastasis;resisting cell death INPUT: Ataxia-telangiectasia mutated ( ATM ) is a high molecular weight protein serine/threonine kinase that plays a central role in the maintenance of genomic integrity by activating cell cycle checkpoints and promoting repair of DNA double-strand breaks . Little is known about the regulatory mechanisms for ATM expression itself . MicroRNAs are naturally existing regulators that modulate gene expression in a sequence-specific manner . Here , we show that a human microRNA , miR-421 , suppresses ATM expression by targeting the 3'-untranslated region ( 3'UTR ) of ATM transcripts . Ectopic expression of miR-421 resulted in S-phase cell cycle checkpoint changes and an increased sensitivity to ionizing radiation , creating a cellular phenotype similar to that of cells derived from ataxia-telangiectasia ( A-T ) patients . Blocking the interaction between miR-421 and ATM 3'UTR with an antisense morpholino oligonucleotide rescued the defective phenotype caused by miR-421 overexpression , indicating that ATM mediates the effect of miR-421 on cell cycle checkpoint and radiosensitivity . Overexpression of the N-Myc transcription factor , an oncogene frequently amplified in neuroblastoma , induced miR-421 expression , which , in turn , down-regulated ATM expression , establishing a linear signaling pathway that may contribute to N-Myc-induced tumorigenesis in neuroblastoma . Taken together , our findings implicate a previously undescribed regulatory mechanism for ATM expression and ATM-dependent DNA damage response and provide several potential targets for treating neuroblastoma and perhaps A-T . OUTPUT:

evading growth suppressors;genomic instability and mutation

HoC_dynamic_5_shot91

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: ATM(Tel1) and ATR(Rad3) checkpoint kinases phosphorylate the C-terminus of histone H2AX ( H2A in yeasts ) in chromatin flanking DNA damage , establishing a recruitment platform for checkpoint and repair proteins . Phospho-H2A/X ( gammaH2A/X)-binding proteins at double-strand breaks ( DSBs ) have been characterized , but those required for replication stress responses are unknown . Here , we present genetic , biochemical , small angle X-ray scattering ( SAXS ) , and X-ray structural studies of the Schizosaccharomyces pombe Brc1 , a 6-BRCT-domain protein that is structurally related to Saccharomyces cerevisiae Rtt107 and mammalian PTIP . Brc1 binds gammaH2A to form spontaneous and DNA damage-induced nuclear foci . Spontaneous Brc1 foci colocalize with ribosomal DNA repeats , a region prone to fork pausing and genomic instability , whereas DNA damage-induced Brc1 foci colocalize with DSB response factors. gammaH2A binding is critical for Brc1 function . The 1.45 A resolution crystal structure of Brc1-gammaH2A complex shows how variable BRCT insertion loops sculpt tandem-BRCT phosphoprotein-binding pockets to facilitate unique phosphoprotein-interaction specificities , and unveils an acidic DNA-mimicking Brc1 surface . From these results , Brc1 docking to gammaH2A emerges as a critical chromatin-specific response to replication-associated DNA damage . OUTPUT: genomic instability and mutation INPUT: DNA damage signaling and repair take place in a chromatin context . Consequently , chromatin-modifying enzymes , including adenosine triphosphate-dependent chromatin remodeling enzymes , play an important role in the management of DNA double-strand breaks ( DSBs ) . Here , we show that the p400 ATPase is required for DNA repair by homologous recombination ( HR ) . Indeed , although p400 is not required for DNA damage signaling , DNA DSB repair is defective in the absence of p400 . We demonstrate that p400 is important for HR-dependent processes , such as recruitment of Rad51 to DSB ( a key component of HR ) , homology-directed repair , and survival after DNA damage . Strikingly , p400 and Rad51 are present in the same complex and both favor chromatin remodeling around DSBs . Altogether , our data provide a direct molecular link between Rad51 and a chromatin remodeling enzyme involved in chromatin decompaction around DNA DSBs . OUTPUT: genomic instability and mutation INPUT: The ATM/ATR DNA damage checkpoint functions in the maintenance of genetic stability and some missense variants of the ATM gene have been shown to confer a moderate increased risk of prostate cancer . However , whether inactivation of this checkpoint contributes directly to prostate specific cancer predisposition is still unknown . Here , we show that exposure of non-malignant prostate epithelial cells ( HPr-1AR ) to androgen led to activation of the ATM/ATR DNA damage response and induction of cellular senescence . Notably , knockdown of the ATM gene expression in HPr-1AR cells can promote androgen-induced TMPRSS2 : ERG rearrangement , a prostate-specific chromosome translocation frequently found in prostate cancer cells . Intriguingly , unlike the non-malignant prostate epithelial cells , the ATM/ATR DNA damage checkpoint appears to be defective in prostate cancer cells , since androgen treatment only induced a partial activation of the DNA damage response . This mechanism appears to preserve androgen induced autophosphorylation of ATM and phosphorylation of H2AX , lesion processing and repair pathway yet restrain ATM/CHK1/CHK2 and p53 signaling pathway . Our findings demonstrate that ATM/ATR inactivation is a crucial step in promoting androgen-induced genomic instability and prostate carcinogenesis . OUTPUT: enabling replicative immortality;genomic instability and mutation INPUT: DNA damage checkpoints arrest cell cycle progression to facilitate DNA repair . The ability to survive genotoxic insults depends not only on the initiation of cell cycle checkpoints but also on checkpoint maintenance . While activation of DNA damage checkpoints has been studied extensively , molecular mechanisms involved in sustaining and ultimately inactivating cell cycle checkpoints are largely unknown . Here , we explored feedback mechanisms that control the maintenance and termination of checkpoint function by computationally identifying an evolutionary conserved mitotic phosphorylation network within the DNA damage response . We demonstrate that the non-enzymatic checkpoint adaptor protein 53BP1 is an in vivo target of the cell cycle kinases Cyclin-dependent kinase-1 and Polo-like kinase-1 ( Plk1 ) . We show that Plk1 binds 53BP1 during mitosis and that this interaction is required for proper inactivation of the DNA damage checkpoint. 53BP1 mutants that are unable to bind Plk1 fail to restart the cell cycle after ionizing radiation-mediated cell cycle arrest . Importantly , we show that Plk1 also phosphorylates the 53BP1-binding checkpoint kinase Chk2 to inactivate its FHA domain and inhibit its kinase activity in mammalian cells . Thus , a mitotic kinase-mediated negative feedback loop regulates the ATM-Chk2 branch of the DNA damage signaling network by phosphorylating conserved sites in 53BP1 and Chk2 to inactivate checkpoint signaling and control checkpoint duration . OUTPUT: evading growth suppressors INPUT: BRIT1 protein ( also known as MCPH1 ) contains 3 BRCT domains which are conserved in BRCA1 , BRCA2 , and other important molecules involved in DNA damage signaling , DNA repair , and tumor suppression . BRIT1 mutations or aberrant expression are found in primary microcephaly patients as well as in cancer patients . Recent in vitro studies suggest that BRIT1/MCPH1 functions as a novel key regulator in the DNA damage response pathways . To investigate its physiological role and dissect the underlying mechanisms , we generated BRIT1(-/-) mice and identified its essential roles in mitotic and meiotic recombination DNA repair and in maintaining genomic stability . Both BRIT1(-/-) mice and mouse embryonic fibroblasts ( MEFs ) were hypersensitive to gamma-irradiation . BRIT1(-/-) MEFs and T lymphocytes exhibited severe chromatid breaks and reduced RAD51 foci formation after irradiation . Notably , BRIT1(-/-) mice were infertile and meiotic homologous recombination was impaired . BRIT1-deficient spermatocytes exhibited a failure of chromosomal synapsis , and meiosis was arrested at late zygotene of prophase I accompanied by apoptosis . In mutant spermatocytes , DNA double-strand breaks ( DSBs ) were formed , but localization of RAD51 or BRCA2 to meiotic chromosomes was severely impaired . In addition , we found that BRIT1 could bind to RAD51/BRCA2 complexes and that , in the absence of BRIT1 , recruitment of RAD51 and BRCA2 to chromatin was reduced while their protein levels were not altered , indicating that BRIT1 is involved in mediating recruitment of RAD51/BRCA2 to the damage site . Collectively , our BRIT1-null mouse model demonstrates that BRIT1 is essential for maintaining genomic stability in vivo to protect the hosts from both programmed and irradiation-induced DNA damages , and its depletion causes a failure in both mitotic and meiotic recombination DNA repair via impairing RAD51/BRCA2's function and as a result leads to infertility and genomic instability in mice . OUTPUT: genomic instability and mutation INPUT: DNA double-strand breaks ( DSBs ) trigger ATM ( ataxia telangiectasia mutated ) signalling and elicit genomic rearrangements and chromosomal fragmentation if misrepaired or unrepaired . Although most DSB repair is ATM-independent , approximately 15% of ionizing radiation ( IR)-induced breaks persist in the absence of ATM-signalling. 53BP1 ( p53-binding protein 1 ) facilitates ATM-dependent DSB repair but is largely dispensable for ATM activation or checkpoint arrest . ATM promotes DSB repair within heterochromatin by phosphorylating KAP-1 ( KRAB-associated protein 1 , also known as TIF1beta , TRIM28 or KRIP-1 ; ref. 2 ) . Here , we show that the ATM signalling mediator proteins MDC1 , RNF8 , RNF168 and 53BP1 are also required for heterochromatic DSB repair . Although KAP-1 phosphorylation is critical for 53BP1-mediated repair , overall phosphorylated KAP-1 ( pKAP-1 ) levels are only modestly affected by 53BP1 loss. pKAP-1 is transiently pan-nuclear but also forms foci overlapping with gammaH2AX in heterochromatin . Cells that do not form 53BP1 foci , including human RIDDLE ( radiosensitivity , immunodeficiency , dysmorphic features and learning difficulties ) syndrome cells , fail to form pKAP-1 foci. 53BP1 amplifies Mre11-NBS1 accumulation at late-repairing DSBs , concentrating active ATM and leading to robust , localized pKAP-1 . We propose that ionizing-radiation induced foci ( IRIF ) spatially concentrate ATM activity to promote localized alterations in regions of chromatin otherwise inhibitory to repair . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot92

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Diphenyl ditelluride ( DPDT ) is a potential prototype for the development of novel biologically active molecules . Thus , it is important to evaluate the toxic effects of this compound . In the present study , we evaluated the cytotoxic , genotoxic and mutagenic properties of DPDT in Chinese hamster fibroblast ( V79 ) cells , in strains of the yeast Saccharomyces cerevisiae both proficient and deficient in several DNA repair pathways and in Salmonella typhimurium . DPDT induced frameshift mutations in both S.typhimurium and a haploid wild-type strain of S.cerevisiae . Mutants of S.cerevisiae defective in base excision repair and recombinational repair were more sensitive to DPDT . The results of a lactate dehydrogenase leakage assay suggest that DPDT is cytotoxic to V79 cells . At cytotoxic concentrations , this compound increased thiobarbituric reactive species levels and decreased the glutathione:GSSH ratio in yeast and V79 cells . DPDT generated single- and double-strand DNA breaks in V79 cells , both with and without metabolic activation , as revealed by alkaline and neutral comet assays . Moreover , an induction of oxidative DNA base damage was indicated by a modified comet assay using formamidopyrimidine DNA glycosylase and endonuclease III . Treatment with DPDT also induced micronucleus formation in V79 cells . Pre-incubation with N-acetylcysteine reduced DPDT's oxidative , genotoxic and mutagenic effects in yeast and V79 cells . Our results suggest that the toxic and mutagenic properties of DPDT may stem from its ability to disturb the redox balance of the cell , which leads to oxidative stress and the induction of DNA damage . OUTPUT: genomic instability and mutation INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: In response to genotoxic stress , a transient arrest in cell-cycle progression enforced by the DNA-damage checkpoint ( DDC ) signalling pathway positively contributes to genome maintenance . Because hyperactivated DDC signalling can lead to a persistent and detrimental cell-cycle arrest , cells must tightly regulate the activity of the kinases involved in this pathway . Despite their importance , the mechanisms for monitoring and modulating DDC signalling are not fully understood . Here we show that the DNA-repair scaffolding proteins Slx4 and Rtt107 prevent the aberrant hyperactivation of DDC signalling by lesions that are generated during DNA replication in Saccharomyces cerevisiae . On replication stress , cells lacking Slx4 or Rtt107 show hyperactivation of the downstream DDC kinase Rad53 , whereas activation of the upstream DDC kinase Mec1 remains normal . An Slx4-Rtt107 complex counteracts the checkpoint adaptor Rad9 by physically interacting with Dpb11 and phosphorylated histone H2A , two positive regulators of Rad9-dependent Rad53 activation . A decrease in DDC signalling results from hypomorphic mutations in RAD53 and H2A and rescues the hypersensitivity to replication stress of cells lacking Slx4 or Rtt107 . We propose that the Slx4-Rtt107 complex modulates Rad53 activation by a competition-based mechanism that balances the engagement of Rad9 at replication-induced lesions . Our findings show that DDC signalling is monitored and modulated through the direct action of DNA-repair factors . OUTPUT: genomic instability and mutation INPUT: Onconase ( Onc ) is an amphibian ribonuclease of the pancreatic RNase family that is cytostatic and cytotoxic to several tumor lines . It also shows anti-tumor activity in mouse tumor models and is currently in phase III clinical trials . In animal tests and clinical trials Onc shows lesser toxicity and fewer side effects compared to most chemotherapeutic drugs . Intriguingly , repeated infusions of this protein do not cause apparent immunological reactions in patients . The aim of the present study was to investigate sensitivity to Onc of human lymphocytes during their mitogenic stimulation in response to the polyvalent mitogen phytohemagglutinin ( PHA ) , and in mixed allogeneic lymphocyte cultures . Unexpectedly , we observed that frequency of cells undergoing activation-induced apoptosis was markedly increased in all cultures containing Onc . Apoptosis was measured by flow cytometry using markers that detect activation of caspases , the in situ presence of DNA strand breaks , and loss of fragmented DNA ( 'sub-G1 ' cell subpopulation ) . The enhancement of frequency of activation-induced apoptosis ( up to 244% ) was observed at 4.2-83 nM Onc concentration , which is at least an order magnitude lower than its minimal concentration reported to affect proliferation or induce apoptosis of leukemic and solid tumor cell lines . The cell cycle progression of lymphocytes that responded to PHA mitogenically was not affected at 8.3 or 83 nM Onc concentration . Because activation-induced apoptosis is the key mechanism regulating several in vivo immunological functions including induction of tolerance , the observed effects of Onc may explain the apparent lack of immune reactions to this protein in treated patients . The propensity of Onc to potentiate the activation-induced apoptosis suggests that this drug may have clinical utility as immunomodulating agent , e.g. , to suppress transplant rejection or treat autoimmune diseases . OUTPUT: resisting cell death;avoiding immune destruction INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Mutations in the gene encoding cytosolic Cu,Zn-superoxide dismutase ( SOD1 ) have been linked to familial amyotrophic lateral sclerosis ( FALS ) . However the molecular mechanisms of motor neuron death are multi-factorial and remain unclear . Here we examined DNA damage , p53 activity and apoptosis in SH-SY5Y human neuroblastoma cells transfected to achieve low-level expression of either wild-type or mutant Gly(93)-->Ala ( G93A ) SOD1 , typical of FALS . DNA damage was investigated by evaluating the levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine ( 8-oxodGuo ) and DNA strand breaks . Significantly higher levels of DNA damage , increased p53 activity , and a greater percentage of apoptotic cells were observed in SH-SY5Y cells transfected with G93A SOD1 when compared to cells overexpressing wild-type SOD1 and untransfected cells . Western blot , FACS , and confocal microscopy analysis demonstrated that G93A SOD1 is present in the nucleus in association with DNA . Nuclear G93A SOD1 has identical superoxide dismutase activity but displays increased peroxidase activity when compared to wild-type SOD1 . These results indicate that the G93A mutant SOD1 association with DNA might induce DNA damage and trigger the apoptotic response by activating p53 . This toxic activity of mutant SOD1 in the nucleus may play an important role in the complex mechanisms associated with motor neuron death observed in ALS pathogenesis . OUTPUT:

genomic instability and mutation;resisting cell death

HoC_dynamic_5_shot93

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Genomic instability drives tumorigenesis , but how it is initiated in sporadic neoplasias is unknown . In early preneoplasias , alterations at chromosome fragile sites arise due to DNA replication stress . A frequent , perhaps earliest , genetic alteration in preneoplasias is deletion within the fragile FRA3B/FHIT locus , leading to loss of Fhit protein expression . Because common chromosome fragile sites are exquisitely sensitive to replication stress , it has been proposed that their clonal alterations in cancer cells are due to stress sensitivity rather than to a selective advantage imparted by loss of expression of fragile gene products . Here , we show in normal , transformed , and cancer-derived cell lines that Fhit-depletion causes replication stress-induced DNA double-strand breaks . Using DNA combing , we observed a defect in replication fork progression in Fhit-deficient cells that stemmed primarily from fork stalling and collapse . The likely mechanism for the role of Fhit in replication fork progression is through regulation of Thymidine kinase 1 expression and thymidine triphosphate pool levels ; notably , restoration of nucleotide balance rescued DNA replication defects and suppressed DNA breakage in Fhit-deficient cells . Depletion of Fhit did not activate the DNA damage response nor cause cell cycle arrest , allowing continued cell proliferation and ongoing chromosomal instability . This finding was in accord with in vivo studies , as Fhit knockout mouse tissue showed no evidence of cell cycle arrest or senescence yet exhibited numerous somatic DNA copy number aberrations at replication stress-sensitive loci . Furthermore , cells established from Fhit knockout tissue showed rapid immortalization and selection of DNA deletions and amplifications , including amplification of the Mdm2 gene , suggesting that Fhit loss-induced genome instability facilitates transformation . We propose that loss of Fhit expression in precancerous lesions is the first step in the initiation of genomic instability , linking alterations at common fragile sites to the origin of genome instability . OUTPUT: genomic instability and mutation;sustaining proliferative signaling;enabling replicative immortality INPUT: Tumor cells are elusive targets for immunotherapy due to their heterogeneity and genetic instability . Here we describe a novel , oral DNA vaccine that targets stable , proliferating endothelial cells in the tumor vasculature rather than tumor cells . Targeting occurs through upregulated vascular-endothelial growth factor receptor 2 ( FLK-1 ) of proliferating endothelial cells in the tumor vasculature . This vaccine effectively protected mice from lethal challenges with melanoma , colon carcinoma and lung carcinoma cells and reduced growth of established metastases in a therapeutic setting . CTL-mediated killing of endothelial cells indicated breaking of peripheral immune tolerance against this self antigen , resulting in markedly reduced dissemination of spontaneous and experimental pulmonary metastases . Angiogenesis in the tumor vasculature was suppressed without impairment of fertility , neuromuscular performance or hematopoiesis , albeit with a slight delay in wound healing . Our strategy circumvents problems in targeting of genetically unstable tumor cells . This approach may provide a new strategy for the rational design of cancer therapies . OUTPUT: activating invasion and metastasis;avoiding immune destruction;inducing angiogenesis INPUT: Progression to malignancy requires that cells overcome senescence and switch to an immortal phenotype . Thus , exploring the genetic and epigenetic changes that occur during senescence/immortalization may help elucidate crucial events that lead to cell transformation . In the present study , we have globally profiled DNA methylation in relation to gene expression in primary , senescent and immortalized mouse embryonic fibroblasts . Using a high-resolution genome-wide mapping technique , followed by extensive locus-specific validation assays , we have identified 24 CpG islands that display significantly higher levels of CpG methylation in immortalized cell lines as compared to primary murine fibroblasts . Several of these hypermethylated CpG islands are associated with genes involved in the MEK-ERK pathway , one of the most frequently disrupted pathways in cancer . Approximately half of the hypermethylated targets are developmental regulators , and bind to the repressive Polycomb group ( PcG ) proteins , often in the context of bivalent chromatin in mouse embryonic stem cells . Because PcG-associated aberrant DNA methylation is a hallmark of several human malignancies , our methylation data suggest that epigenetic reprogramming of pluripotency genes may initiate cell immortalization . Consistent with methylome alterations , global gene expression analysis reveals that the vast majority of genes dysregulated during cell immortalization belongs to gene families that converge into the MEK-ERK pathway . Additionally , several dysregulated members of the MAP kinase network show concomitant hypermethylation of CpG islands . Unlocking alternative epigenetic routes for cell immortalization will be paramount for understanding crucial events leading to cell transformation . Unlike genetic alterations , epigenetic changes are reversible events , and as such , can be amenable to pharmacological interventions , which makes them appealing targets for cancer therapy when genetic approaches prove inadequate . OUTPUT: enabling replicative immortality INPUT: Human carcinomas are defined by recurrent chromosomal aneuploidies , which result in a tissue-specific distribution of genomic imbalances . In order to develop models for these genome mutations and to determine their role in tumorigenesis , we generated 45 spontaneously transformed murine cell lines from normal epithelial cells derived from bladder , cervix , colon , kidney , lung , and mammary gland . Phenotypic changes , chromosomal aberrations , centrosome number , and telomerase activity were assayed in control uncultured cells and in three subsequent stages of transformation . Supernumerary centrosomes , binucleate cells , and tetraploidy were observed as early as 48 hr after explantation . In addition , telomerase activity increased throughout progression . Live-cell imaging revealed that failure of cytokinesis , not cell fusion , promoted genome duplication . Spectral karyotyping demonstrated that aneuploidy preceded immortalization , consisting predominantly of whole chromosome losses ( 4 , 9 , 12 , 13 , 16 , and Y ) and gains ( 1 , 10 , 15 , and 19 ) . After transformation , focal amplifications of the oncogenes Myc and Mdm2 were frequently detected . Fifty percent of the transformed lines resulted in tumors on injection into immunocompromised mice . The phenotypic and genomic alterations observed in spontaneously transformed murine epithelial cells recapitulated the aberration pattern observed during human carcinogenesis . The dominant aberration of these cell lines was the presence of specific chromosomal aneuploidies . We propose that our newly derived cancer models will be useful tools to dissect the sequential steps of genome mutations during malignant transformation , and also to identify cancer-specific genes , signaling pathways , and the role of chromosomal instability in this process . OUTPUT: enabling replicative immortality;genomic instability and mutation INPUT: The identification of molecular markers related to critical biological processes during carcinogenesis may aid in the evaluation of carcinogenic potentials of chemicals and chemical mixtures . Work from our laboratory demonstrated that a single treatment with N-methyl-N'-nitro-N-nitrosoguanidine ( MNNG ) enhanced spontaneous malignant transformation of the human keratinocyte cell line RHEK-1 . In contrast , chronic low-level exposure of cells to arsenic alone or in a mixture containing arsenic , cadmium , chromium , and lead inhibited malignant conversion . To identify changes in gene expression that influence these different outcomes , cDNA microarray technology was used . Analysis of multiple human arrays in MNNG-transformed RHEK-1 cells , designated OM3 , and those treated with arsenic or the arsenic-containing metal mixture showed unique patterns of gene expression . Genes that were overexpressed in OM3 included oncogenes , cell cycle regulators , and those involved in signal transduction , whereas genes for DNA repair enzymes and inhibitors of transformation and metastasis were suppressed . In arsenic-treated cells , multiple DNA repair proteins were overexpressed . Mixture-treated cells showed increased expression of a variety of genes including metallothioneins and integrin 4 . These cells showed decreased expression of oncogenes , DNA repair proteins , and genes involved in the mitogen-activated protein kinase pathway . For comparison we are currently analyzing gene expression changes in RHEK-1 cells transformed by other means . The goal of these studies is to identify common batteries of genes affected by chemical modulators of the carcinogenic process . Mechanistic studies may allow us to correlate alterations in their expression with sequential stages in the carcinogenic process and may aid in the risk assessment of other xenobiotics . OUTPUT: activating invasion and metastasis;evading growth suppressors;genomic instability and mutation;sustaining proliferative signaling INPUT: During tumorigenesis , cells acquire immortality in association with the development of genomic instability . However , it is still elusive how genomic instability spontaneously generates during the process of tumorigenesis . Here , we show that precancerous DNA lesions induced by oncogene acceleration , which induce situations identical to the initial stages of cancer development , trigger tetraploidy/aneuploidy generation in association with mitotic aberration . Although oncogene acceleration primarily induces DNA replication stress and the resulting lesions in the S phase , these lesions are carried over into the M phase and cause cytokinesis failure and genomic instability . Unlike directly induced DNA double-strand breaks , DNA replication stress-associated lesions are cryptogenic and pass through cell-cycle checkpoints due to limited and ineffective activation of checkpoint factors . Furthermore , since damaged M-phase cells still progress in mitotic steps , these cells result in chromosomal mis-segregation , cytokinesis failure and the resulting tetraploidy generation . Thus , our results reveal a process of genomic instability generation triggered by precancerous DNA replication stress . OUTPUT:

evading growth suppressors;genomic instability and mutation

HoC_dynamic_5_shot94

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Cancer cells constantly adapt to oxidative phosphorylation ( OXPHOS ) suppression resulting from hypoxia or mitochondria defects . Under the OXPHOS suppression , AMP-activated protein kinase ( AMPK ) regulates global metabolism adjustments , but its activation has been found to be transient . Whether cells can maintain cellular ATP homeostasis and survive beyond the transient AMPK activation is not known . Here , we study the bioenergetic adaptation to the OXPHOS inhibitor oligomycin in a group of cancer cells . We found that oligomycin at 100 ng/ml completely inhibits OXPHOS activity in 1 h and induces various levels of glycolysis gains by 6 h , from which we calculate the bioenergetic organizations of cancer cells . In glycolysis-dominant cells , oligomycin does not induce much energy stress as measured by glycolysis acceleration , ATP imbalance , AMPK activation , AMPK substrate acetyl-CoA carboxylase phosphorylation at Ser(79) , and cell growth inhibition . In OXPHOS-dependent LKB1 wild type cells , oligomycin induces 5-8% ATP drops and transient AMPK activation during the initial 1-2 h . After AMPK activation is completed , oligomycin-induced increase of acetyl-CoA carboxylase phosphorylation at Ser(79) is still detected , and cellular ATP is back at preoligomycin treatment levels by sustained elevation of glycolysis . Cell growth , however , is inhibited without an increase in cell death and alteration in cell cycle distribution . In OXPHOS-dependent LKB1-null cells , no AMPK activation by oligomycin is detected , yet cells still show a similar adaptation . We also demonstrate that the adaptation to oligomycin does not invoke activation of hypoxia-induced factor . Our data suggest that cancer cells may grow and survive persistent OXPHOS suppression through an as yet unidentified regulatory mechanism . OUTPUT: cellular energetics INPUT: The high glucose consumption of tumor cells even in an oxygen-rich environment , referred to as the Warburg effect , has been noted as a nearly universal biochemical characteristic of cancer cells . Targeting the glycolysis pathway has been explored as an anti-cancer therapeutic strategy to eradicate cancer based on this fundamental biochemical property of cancer cells . Oncoproteins such as Akt and c-Myc regulate cell metabolism . Accumulating studies have uncovered various molecular mechanisms by which oncoproteins affect cellular metabolism , raising a concern as to whether targeting glycolysis will be equally effective in treating cancers arising from different oncogenic activities . Here , we established a dual-regulatable FL5.12 pre-B cell line in which myristoylated Akt is expressed under the control of doxycycline , and c-Myc , fused to the hormone-binding domain of the human estrogen receptor , is activated by 4-hydroxytamoxifen . Using this system , we directly compared the effect of these oncoproteins on cell metabolism in an isogenic background . Activation of either Akt or c-Myc leads to the Warburg effect as indicated by increased cellular glucose uptake , glycolysis , and lactate generation . When cells are treated with glycolysis inhibitors , Akt sensitizes cells to apoptosis , whereas c-Myc does not . In contrast , c-Myc but not Akt sensitizes cells to the inhibition of mitochondrial function . This is correlated with enhanced mitochondrial activities in c-Myc cells . Hence , although both Akt and c-Myc promote aerobic glycolysis , they differentially affect mitochondrial functions and render cells susceptible to the perturbation of cellular metabolic programs . OUTPUT: cellular energetics INPUT: Cancer cells upregulate glycolysis , increasing glucose uptake to meet energy needs . A small fraction of a cell's glucose enters the hexosamine biosynthetic pathway ( HBP ) , which regulates levels of O-linked beta-N-acetylglucosamine ( O-GlcNAc ) , a carbohydrate posttranslational modification of diverse nuclear and cytosolic proteins . We discovered that breast cancer cells upregulate the HBP , including increased O-GlcNAcation and elevated expression of O-GlcNAc transferase ( OGT ) , which is the enzyme catalyzing the addition of O-GlcNAc to proteins . Reduction of O-GlcNAcation through RNA interference of OGT in breast cancer cells leads to inhibition of tumor growth both in vitro and in vivo and is associated with decreased cell-cycle progression and increased expression of the cell-cycle inhibitor p27(Kip1) . Elevation of p27(Kip1) was associated with decreased expression and activity of the oncogenic transcription factor FoxM1 , a known regulator of p27(Kip1) stability through transcriptional control of Skp2 . Reducing O-GlcNAc levels in breast cancer cells decreased levels of FoxM1 protein and caused a decrease in multiple FoxM1-specific targets , including Skp2 . Moreover , reducing O-GlcNAcation decreased cancer cell invasion and was associated with the downregulation of matrix metalloproteinase-2 , a known FoxM1 target . Finally , pharmacological inhibition of OGT in breast cancer cells had similar anti-growth and anti-invasion effects . These findings identify O-GlcNAc as a novel mechanism through which alterations in glucose metabolism regulate cancer growth and invasion and suggest that OGT may represent novel therapeutic targets for breast cancer . OUTPUT: evading growth suppressors;cellular energetics INPUT: Cancer cells typically display altered glucose metabolism characterized by a preference of aerobic glycolysis , known as the Warburg effect , which facilitates cell proliferation . Hypoxia-inducible factor ( HIF ) and oncoprotein Myc are two prominent transcription factors that drive glycolysis . Previously , we reported that the estrogen-related receptors ( ERRs ) act as cofactors of HIF and enhance HIF-dependent transcription of glycolytic genes under hypoxia . ERRs are orphan nuclear receptors and key regulators of energy metabolism by orchestrating mitochondrial biogenesis , fatty acid oxidation ( FAO ) and oxidative phosphorylation . Here , we show that ERRs also stimulate glycolysis under normoxia . ERRs directly bind to and activate promoters of many genes encoding glycolytic enzymes , and the ERR-binding sites in such promoters are essential for ERR-mediated transcriptional activation . ERRs interact with Myc , and the two factors synergistically activate transcription of glycolytic genes . Furthermore , overexpression of ERRs increases glycolytic gene expression and lactate production . Conversely , depletion of ERRs in cancer cells reduces expression of glycolytic genes and glucose uptake , resulting in decreased aerobic glycolysis and cell growth . Taken together , these results suggest that ERRs are important transcriptional activators of the glycolytic pathway and contribute to the Warburg effect in cancer cells . OUTPUT: cellular energetics;sustaining proliferative signaling INPUT: Mitochondrial biogenesis , which depends on nuclear as well as mitochondrial genes , occurs in response to increased cellular ATP demand . The nuclear transcriptional factors , estrogen-related receptor alpha ( ERRalpha ) and nuclear respiratory factors 1 and 2 , are associated with the coordination of the transcriptional machinery governing mitochondrial biogenesis , whereas coactivators of the peroxisome proliferator-activated receptor gamma coactivator-1 ( PGC-1 ) family serve as mediators between the environment and this machinery . In the context of proliferating cells , PGC-1-related coactivator ( PRC ) is a member of the PGC-1 family , which is known to act in partnership with nuclear respiratory factors , but no functional interference between PRC and ERRalpha has been described so far . We explored three thyroid cell lines , FTC-133 , XTC.UC1 and RO 82 W-1 , each characterized by a different mitochondrial content , and studied their behavior towards PRC and ERRalpha in terms of respiratory efficiency . Overexpression of PRC and ERRalpha led to increased respiratory chain capacity and mitochondrial mass . The inhibition of ERRalpha decreased cell growth and respiratory chain capacity in all three cell lines . However , the inhibition of PRC and ERRalpha produced a greater effect in the oxidative cell model , decreasing the mitochondrial mass and the phosphorylating respiration , whereas the nonphosphorylating respiration remained unchanged . We therefore hypothesize that the ERRalpha-PRC complex plays a role in arresting the cell cycle through the regulation of oxidative phosphorylation in oxidative cells , and through some other pathway in glycolytic cells . OUTPUT: cellular energetics INPUT: p32/gC1qR/C1QBP/HABP1 is a mitochondrial/cell surface protein overexpressed in certain cancer cells . Here we show that knocking down p32 expression in human cancer cells strongly shifts their metabolism from oxidative phosphorylation ( OXPHOS ) to glycolysis . The p32 knockdown cells exhibited reduced synthesis of the mitochondrial-DNA-encoded OXPHOS polypeptides and were less tumorigenic in vivo . Expression of exogenous p32 in the knockdown cells restored the wild-type cellular phenotype and tumorigenicity . Increased glucose consumption and lactate production , known as the Warburg effect , are almost universal hallmarks of solid tumors and are thought to favor tumor growth . However , here we show that a protein regularly overexpressed in some cancers is capable of promoting OXPHOS . Our results indicate that high levels of glycolysis , in the absence of adequate OXPHOS , may not be as beneficial for tumor growth as generally thought and suggest that tumor cells use p32 to regulate the balance between OXPHOS and glycolysis . OUTPUT:

cellular energetics

HoC_dynamic_5_shot95

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: OBJECTIVE To study the clinicopathologic features , immunophenotype and ultrastructural features of sinonasal inflammatory myofibroblastic tumors ( IMT ) . METHODS The clinical and histologic features of 5 cases of sinonasal IMT were reviewed . Immunohistochemical study for vimentin , MSA , SMA , calponin , h-caldesmon , desmin , ALK , fibronectin , CK , S-100 and Ki-67 was carried out . Ultrastructural examination was also performed in two of the cases . RESULTS The patients age ranged from 28 to 62 years ( mean = 43 years ) . The male-to-female ratio was 2:3 . The clinical presentation included nasal obstruction , nasal discharge , nasal bleeding , facial pain , facial swelling , toothache and tear overflow . All of the 5 patients suffered from disease relapses ; and 4 of them had recurrences for more than 5 times . One patient had lymph node metastasis and 3 patients died of the disease . Histologically , the tumor cells were arranged in interlacing fascicles and sometimes haphazard in fashion . They were spindly in shape , cytoplasm eosinophilic with mild nuclear atypia and a low mitotic activity . The intervening stroma was myxoid in appearance accompanied by lymphocyte and plasma cell infiltration , abundant blood vessels and focal collagenized areas . In 3 of the recurrent cases , the tumor cells displayed increased nuclear atypia and mitotic activity ( average about 5 to 6 per 10 high-power fields ) , accompanied by patchy necrosis , less inflammatory cell infiltration and focal sarcomatous changes . Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin . SMA , MSA , calponin and fibronectin were variably expressed . Desmin was weakly positive in 1 case . The staining for h-caldesmon , ALK , S-100 and CK was negative . The Ki-67 proliferation index increased with tumor recurrences . Electron microscopy revealed abundant rough endoplasmic reticulum and dense body formation in the cytoplasm . There were an increased amount of collagen fibers in the stroma . CONCLUSIONS IMT rarely occurs in nasal cavity and paranasal sinuses . The tumor is prone to local invasion and recurrences , with subsequent progression to frank malignancy and distant metastasis , resulting in high mortality and poor prognosis . Complete surgical resection remains the main modality of treatment . OUTPUT: activating invasion and metastasis;resisting cell death;tumor promoting inflammation;sustaining proliferative signaling INPUT: Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur . The single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases . Thus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA . To assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast . The reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37°C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection . This exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang . We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G . APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes . We then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake . Sulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes . Furthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase . This intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol ζ , during repair synthesis . Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately . OUTPUT: genomic instability and mutation INPUT: Malignant mesothelioma ( MM ) still remains a therapeutic and diagnostic problem to which new therapeutic perspectives are being continuously tried and tested . Three different primary cultures ( MMGe-1 , MES MM 98 , and MES 1 ) and one immortalized cell line ( MSTO 211 H ) of human MM were studied in order to evaluate the HER-2/neu expression . Three out of four cell lines showed a different level of c-erbB-2 expression , the highest being detected on the MSTO 211 H cell line ( fibroblastic phenotype ) , whereas MMGe-1 resulted negative . The effect of the anti-HER-2/neu antibody ( Trastuzumab ) alone , and in combination with cisplatin ( CDDP ) at different doses ( ranging from 0.1 to 100 microg/ml ) , was studied on all the c-erB-2 positive cell lines . Trastuzumab was able to inhibit cell proliferation in a time-dependent manner , with growth inhibition also obtained at low concentrations ( 0.1-1 microg/ml ) . Combined treatment with Trastuzumab ( 10 microg/ml ) and CDDP ( 1 microg/ml ) showed synergism . Our results were encouraging , and suggest a rationale for further investigations in a clinical setting . OUTPUT: sustaining proliferative signaling INPUT: Systemic oxidative stress is associated with a wide range of pathological conditions . Oxidative DNA damage is frequently measured in circulating lymphocytes . Mitochondrial DNA ( mtDNA ) is known to be more sensitive to oxidative damage than nuclear DNA but is rarely used for direct measurement of DNA damage in clinical studies . Based on the supercoiling-sensitive real-time PCR method , we propose a new approach for the noninvasive monitoring of systemic oxidative stress by quantifying the mtDNA structural damage and copy number change in isolated lymphocytes in a single test . We show that lymphocytes have significantly less mtDNA content and relatively lower baseline levels of damage than cancer cell lines . In an ex vivo challenge experiment , we demonstrate , for the first time , that exogenous H2O2 induces a significant increase in mtDNA damage in lymphocytes from healthy individuals , but no repair activity is observed after 1 h recovery . We further demonstrate that whole blood may serve as a convenient alternative to the isolated lymphocytes in mtDNA analysis . Thus , the blood analysis with the multiple mtDNA end-points proposed in the current study may provide a simple and sensitive test to interrogate the nature and extent of systemic oxidative stress for a broad spectrum of clinical investigations . OUTPUT: genomic instability and mutation;tumor promoting inflammation INPUT: BACKGROUND Engineered zinc-finger nucleases ( ZFN ) represented an innovative method for the genome manipulation in vertebrates . ZFN introduced targeted DNA double strand breaks ( DSB ) and initiated non-homologous end joining ( NHEJ ) after pronuclear or cytoplasmatic microinjection into zygotes . Resulting frame shift mutations led to functional gene ablations in zebra fish , mice , pigs and also in laboratory rats . Therefore , we targeted the rat Rag1 gene essential for the V(D)J recombination within the immunoglobulin production process and for the differentiation of mature B and T lymphocytes to generate an immunodeficient rat model in the LEW/Ztm strain . RESULTS After microinjection of Rag1 specific ZFN mRNAs in 623 zygotes of inbred LEW/Ztm rats 59 offspring were born from which one carried a 4 bp deletion . This frame shift mutation led to a premature stop codon and a subsequently truncated Rag1 protein confirmed by the loss of the full-length protein in Western Blot analysis . Truncation of the Rag1 protein was characterized by the complete depletion of mature B cells . The remaining T cell population contained mature CD4+/CD3+/TCRαβ+ as well as CD8+/CD3+/TCRαβ+ positive lymphocytes accompanied by a compensatory increase of natural killer cells in the peripheral blood . Reduction of T cell development in Rag1 mutant rats was associated with a hypoplastic thymus that lacked follicular structures . Histological evaluation also revealed the near-complete absence of lymphocytes in spleen and lymph nodes in the immunodeficient Rag1 mutant rat . CONCLUSION The Rag1 mutant rat will serve as an important model for transplantation studies . Furthermore , it may be used as a model for reconstitution experiments related to the immune system , particularly with respect to different populations of human lymphocytes , natural killer cells and autoimmune phenomena . OUTPUT: genomic instability and mutation;avoiding immune destruction INPUT: The contribution of a type II restriction-modification system ( R-M system ) to genome integrity and cell viability was investigated . We established experimental conditions which enabled the achievement of hemimethylated and unmethylated states for the specific bases of the recognition sequences of the host's DNA . To achieve this , we constructed the MboII R-M system containing only one ( i.e . M2.MboII ) out of two functional MboII methyltransferases found in Moraxella bovis . Using the incomplete R-M system we were able to perturb the balance between methylation and restriction in an inducible manner . We demonstrate that upon the SOS-induced DNA repair in the mitomycin C treated cells , restriction significantly reduces cell viability . Similar results for the well-studied wild type EcoRI R-M system , expressed constitutively in Escherichia coli , were obtained . Our data provide further insights into the benefits and disadvantages of maintaining of a type II R-M system , highlighting its impact on host cell fitness . OUTPUT:

genomic instability and mutation

HoC_dynamic_5_shot96

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Aerobic glycolysis and mitochondrial dysfunction are common features of aggressive cancer growth . We observed promoter methylation and loss of expression in neurofilament heavy polypeptide ( NEFH ) in a significant proportion of primary esophageal squamous cell carcinoma ( ESCC ) samples that were of a high tumor grade and advanced stage . RNA interference-mediated knockdown of NEFH accelerated ESCC cell growth in culture and increased tumorigenicity in vivo , whereas forced expression of NEFH significantly inhibited cell growth and colony formation . Loss of NEFH caused up-regulation of pyruvate kinase-M2 type and down-regulation of pyruvate dehydrogenase , via activation of the Akt/beta-catenin pathway , resulting in enhanced aerobic glycolysis and mitochondrial dysfunction . The acceleration of glycolysis and mitochondrial dysfunction in NEFH-knockdown cells was suppressed in the absence of beta-catenin expression , and was decreased by the treatment of 2-Deoxyglucose , a glycolytic inhibitor , or API-2 , an Akt inhibitor . Loss of NEFH activates the Akt/beta-catenin pathway and increases glycolysis and mitochondrial dysfunction . Cancer cells with methylated NEFH can be targeted for destruction with specific inhibitors of deregulated downstream pathways . OUTPUT: cellular energetics INPUT: Although the immense efforts have been made for cancer prevention , early diagnosis , and treatment , cancer morbidity and mortality has not been decreased during last forty years . Especially , lung cancer is top-ranked in cancer-associated human death . Therefore , effective strategy is strongly required for the management of lung cancer . In the present study , we found that novel daphnane diterpenoids , yuanhualine ( YL ) , yuanhuahine ( YH ) and yuanhuagine ( YG ) isolated from the flower of Daphne genkwa ( Thymelaeaceae ) , exhibited potent anti-proliferative activities against human lung A549 cells with the IC50 values of 7.0 , 15.2 and 24.7 nM , respectively . Flow cytometric analysis revealed that the daphnane diterpenoids induced cell-cycle arrest in the G0/G1 as well as G2/M phase in A549 cells . The cell-cycle arrests were well correlated with the expression of checkpoint proteins including the up-regulation of cyclin-dependent kinase inhibitor p21 and p53 and down-regulation of cyclin A , cyclin B1 , cyclin E , cyclin dependent kinase 4 , cdc2 , phosphorylation of Rb and cMyc expression . In the analysis of signal transduction molecules , the daphnane diterpenoids suppressed the activation of Akt , STAT3 and Src in human lung cancer cells . The daphnane diterpenoids also exerted the potent anti-proliferative activity against anticancer-drug resistant cancer cells including gemcitabine-resistant A549 , gefitinib- , erlotinib-resistant H292 cells . Synergistic effects in the growth inhibition were also observed when yuanhualine was combined with gemcitabine , gefitinib or erlotinib in A549 cells . Taken together , these findings suggest that the novel daphnane diterpenoids might provide lead candidates for the development of therapeutic agents for human lung cancers . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: Despite experimental evidence that sulforaphane can exert chemopreventive effects , whether these effects are specific for neoplastic cells is not known . Following our previous demonstration that sulforaphane induces cell cycle arrest and apoptosis in human T lymphoblastoid Jurkat leukemia cells and increases p53 and bax protein expression , we tested sulforaphane on non-transformed phytohemagglutinin-stimulated human lymphocytes . Here , we demonstrate that sulforaphane arrested cell cycle progression in G , phase , through a decrease in the protein expression of cyclin D3 . Moreover , sulforaphane induced apoptosis ( and also necrosis ) , mediated by an increase in the expression of p53 . These findings suggest that sulforaphane is a growth modulator for T cells . Our in vitro evidence that sulforaphane is active and even cytotoxic in normal as well as transformed lymphocytes raises important questions regarding its suitability for cancer chemoprevention . OUTPUT: sustaining proliferative signaling;resisting cell death INPUT: PURPOSE : Acid ceramidase ( AC ) occupies an important place in the control of cancer cell proliferation . We tested the influence of AC inhibition on the effects of PSC 833 , a P-glycoprotein antagonist with potent ceramide-generating capacity , to determine whether AC could be a therapeutic target in pancreatic cancer . METHODS : Ceramide metabolism was followed using ( 3)H-palmitate , and molecular species were determined by mass spectroscopy . Apoptosis was measured by DNA fragmentation , autophagy by acridine orange staining , and cell cycle was assessed by flow cytometry and RB phosphorylation . AC was measured in intact cells using fluorescent substrate . RESULTS : Exposure of human PANC-1 or MIA-PaCa-2 cells to PSC 833 promoted increases in de novo ( dihydro)ceramides , ( dihydro)glucosylceramides , and ( dihydro)sphingomyelins , demarking ceramide generation and robust metabolism . Despite the multifold increases in ( dihydro)ceramide levels , cells were refractory to PSC 833 . However , PSC 833 produced a dose-dependent decrease in DNA synthesis and dose- and time-dependent decreases in RB phosphorylation , consistent with cell cycle arrest as demonstrated at G1 . Cytostatic effects of PSC 833 were converted to cytotoxic end-point by acid ceramidase inhibition . Cytotoxicity was accompanied by formation of acridine orange-stained acidic vesicles and an increase in LC3 expression , indicative of autophagic response . Cell death was not reversed by preexposure to myriocin , which blocks PSC 833-induced ceramide generation . CONCLUSION : Although the role of ceramide in end-point cytotoxicity is unclear , our results suggest that acid ceramidase is a viable target in pancreatic cancer . We propose that AC inhibition will be effective in concert with other anticancer therapies . OUTPUT: sustaining proliferative signaling;resisting cell death INPUT: BACKGROUND AND AIM The importance of glycolysis in cancer cells is well documented . The effects of inhibiting glycolysis using metabolic inhibitors iodoacetate ( IAA ) , an inhibitor of GAPDHase , and 3-bromopyruvate ( 3BP ) , an inhibitor of hexokinase-II , on survival and signaling of pancreatic cancer cells ( Panc-1 ) were investigated . MATERIALS AND METHODS Cellular survival was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ( MTT ) assay . Lactate dehydrogenase ( LDH ) assay was used to analyze the induced necrosis and protein levels were evaluated using Western blot analysis . RESULTS The results show that the inhibitors lowered cellular survival and increased cellular necrosis . Mitogenic signaling pathways were affected by 3BP but not by IAA . CONCLUSION We conclude that there may be a cross-talk between signaling pathways and glycolysis in regulating pancreatic cancer cell survival and signaling . Thus , a combination of agents that inhibit both energy production and cell signaling may provide a novel and effective approach to target pancreatic cancer effectively . OUTPUT: resisting cell death;cellular energetics INPUT: Acidosis commonly observed in solid tumors like pancreatic cancer promotes genetic instability and selection of a more malignant phenotype of cancer cells . Overexpression or activation of integral membrane proteins mediating H+ efflux may contribute to extracellular acidification . Neurotensin ( NT ) induces intracellular alkalinization and stimulates interleukin-8 production in pancreatic cancer cells and , as demonstrated here , the stable NT analog Lys(8)-psi-Lys(9)NT(8-13) enhances the amiloride-sensitive , Na+-dependent transmembrane H+ flux by a factor of 2.05+/-0.28 and 2.69+/-0.07 in BxPC-3 and PANC-1 pancreatic cancer cells , respectively , by phosphorylation of the Na+/H+ exchanger 1 ( NHE1 ) . Human genome-wide gene expression analysis was performed to detect effects of Lys(8)-psi-Lys(9)NT(8-13) on BxPC-3 cells . Results indicated upregulation of genes involved in regulation of NHE1 , hypoxic response and glycolysis in response to Lys(8)-psi-Lys(9)NT(8-13) even under normoxic conditions . Therefore , our findings suggest that growth factors like NT may be implicated in the early progression of pancreatic cancer by localized acidification and induction of an aerobic glycolytic phenotype with higher metastatic potential in small cell aggregates . OUTPUT:

cellular energetics

HoC_dynamic_5_shot97

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells . Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines . Since new compounds with biological activity are needed , the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones , derived from 1-naphthaldehyde and 2-naphthaldehyde , on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells . Based on the results , the most cytotoxic compound ( A1 ) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line ( HT-29 ) . Chalcone A1 significantly reduced the cell viability of K562 , Jurkat , Kasumi , U937 , CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group ( IC(50) values between ∼1.5μM and 40μM ) . It was also cytotoxic to HL-29 cells . To further examine its effect on normal cells , peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound . It has also been incubated with human fibroblasts cultured from bone marrow ( JMA ) . Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells . A1 caused significant cell cycle arrest in all phases according to the cell line , and increased the proportion of cells in the sub G0/G1 phase . To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway , cell morphology was examined using fluorescence microscopy . Cells treated with A1 at IC(50) demonstrated the morphological characteristic of apoptosis , such as chromatin condensation and formation of apoptotic bodies . Apoptosis was confirmed by externalization of phosphatidylserine , which was detected by the Annexin V-FITC method , and by DNA fragmentation . The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy . OUTPUT: evading growth suppressors;sustaining proliferative signaling;resisting cell death INPUT: Colorectal cancer ( CRC ) arises as the consequence of progressive changes from normal epithelial cells through polyp to tumor , and thus is an useful model for studying metabolic shift . In the present study , we studied the metabolomic profiles using high analyte specific gas chromatography/mass spectrometry ( GC/MS ) and liquid chromatography tandem mass spectrometry ( LC/MS/MS ) to attain a systems-level view of the shift in metabolism in cells progressing along the path to CRC . Colonic tissues including tumor , polyps and adjacent matched normal mucosa from 26 patients with sporadic CRC from freshly isolated resections were used for this study . The metabolic profiles were obtained using GC/MS and LC/MS/MS . Our data suggest there was a distinct profile change of a wide range of metabolites from mucosa to tumor tissues . Various amino acids and lipids in the polyps and tumors were elevated , suggesting higher energy needs for increased cellular proliferation . In contrast , significant depletion of glucose and inositol in polyps revealed that glycolysis may be critical in early tumorigenesis . In addition , the accumulation of hypoxanthine and xanthine , and the decrease of uric acid concentration , suggest that the purine biosynthesis pathway could have been substituted by the salvage pathway in CRC . Further , there was a step-wise reduction of deoxycholic acid concentration from mucosa to tumors . It appears that to gain a growth advantage , cancer cells may adopt alternate metabolic pathways in tumorigenesis and this flexibility allows them to adapt and thrive in harsh environment . OUTPUT: cellular energetics INPUT: Spindle cell oncocytoma ( SCO ) of the pituitary gland is a relatively recently established , very rare subtype of adenohypophysis tumours that was introduced as a distinct clinicopathological entity in the fourth edition of WHO classification of the central nervous system tumours ( 2007 ) . It is non-endocrine neoplasm of the anterior pituitary that occurs in adults and usually follows a benign clinical course , corresponding to WHO grade I. Up to now , pituitary SCO have been reported occasionally and only 14 cases of SCO have been documented in the literature . Because of their rarity , the pathogenesis and natural history of these tumours have not been fully characterized . We report two additional cases of SCO occurring in females aged 63 years ( Case 1 ) and 65 years ( Case 2 ) , who presented with pan-hypopituitarism , headache and visual field defect . In both cases , the magnetic resonance imaging showed solid sellar mass of moderate size with suprasellar extension . The clinical and radiological features suggested non-functioning pituitary macroadenomas without evidence of invasive growth . One patient presented with tumour recurrence 3 years after undergoing the previous surgical removal of tumour , which was initially misdiagnosed as schwannoma . The first tumour was removed by transsphenoidal surgery and the second one by frontal craniotomy . Histologically and immunohistochemically , both tumours displayed the features typical for SCO of the pituitary . They were composed of interwoven fascicles of spindle cells exhibiting abundant eosinophilic cytoplasm of oncocytic or granular appearance . Mitoses were rarely observed and necrosis was absent . In one case , the advanced lymphocytic infliltration was observed within neoplastic tissue . The tumour cells exhibited immunoreactivity for S-100 protein , galectin-3 , vimentin and epithelial membrane antigen but they were negative for GFAP , anterior pituitary neuroendocrine markers ( prolactin , growth hormone , TSH , ACTH , FSH , LH ) , chromogranin , synaptophysin , cytokeratin CK ( AE1/AE3 ) , smooth muscle actin , desmin , CD34 and CD68 . MIB1 labeling index did not exceed 10% . Ultrastructurally , the tumour cells were rich in mitochondria with lamellar cristae . Moreover , in Case 2 some tumour cells showed a number of giant mitochondria with severely destructed internal matrix . Spindle cell oncocytoma of the anterior pituitary is often misdiagnosed entity of uncertain histogenesis . It should be considered in the differential diagnosis of various sellar-region lesions of oncocytic morphology . OUTPUT: resisting cell death INPUT: Contact inhibition is a crucial mechanism regulating proliferation in vitro and in vivo . Although it is generally accepted that contact inhibition plays a pivotal role in maintaining tissue homeostasis , the molecular mechanisms of contact inhibition are still not fully understood . FoxM1 is known as a proliferation-associated transcription factor and is upregulated in many cancer types . Vice versa , anti-proliferative signals , such as TGF-β and differentiation signals decrease FoxM1 expression . Here we investigated the role of FoxM1 in contact inhibition in fibroblasts . We show that protein expression of FoxM1 is severely and rapidly downregulated upon contact inhibition , probably by inhibition of ERK activity , which then leads to decreased expression of cyclin A and polo-like kinase 1 . Vice versa , ectopic expression of FoxM1 prevents the decrease in cyclin A and polo-like kinase 1 and causes a two-fold increase in saturation density indicating loss of contact inhibition . Hence , we show that downregulation of FoxM1 is required for contact inhibition by regulating expression of cyclin A and polo-like kinase 1 . OUTPUT: evading growth suppressors INPUT: BACKGROUND Ovarian surface epithelial cells undergo several rounds of division to repair the wound created by follicular rupture at the time of ovulation . This cyclical requirement for cell division , when not interrupted by the long anovulatory rest periods that occur during pregnancy and lactation , may contribute to the development of ovarian cancer . PURPOSE AND METHODS To test this hypothesis , we isolated rat ovarian surface epithelial cells from 10 adult female Fisher rats , initiated two mixed-population and seven clonal cell lines , and repeatedly subcultured these cells in vitro for more than 20 passages . We then tested them for the acquisition of the following four features associated with transformation : 1 ) the loss of contact inhibition , 2 ) the capacity for substrate-independent growth , 3 ) the ability to form tumors when injected subcutaneously and/or intraperitoneally into athymic mice , and 4 ) cytogenetic abnormalities . RESULTS Loss of contact inhibition was observed in all nine late-passage cell lines . Six of the nine late-passage , but none of the early-passage , cell lines tested exhibited a capacity for substrate-independent growth that was augmented in a dose-dependent manner by epidermal growth factor . Two late-passage cell lines ( clone 2 and mixed-population 2 ) generated tumors in athymic BALB/c mice within 3 weeks following subcutaneous injection of 5 x 10(6) cells , whereas similar numbers of early-passage cells from the same cell lines failed to generate palpable tumors . Late-passage clone 7 cells were tumorigenic when 5 x 10(7) cells were injected intraperitoneally . Two of the cell lines analyzed exhibited alterations involving losses of part or all of one member of the chromosome 5 pair . Clone 2 possessed an interstitial deletion , del(5)(q21.3q24) , consistent with the loss of an uncloned putative tumor suppressor gene at 5q22q23 previously reported to reside near the loci for the interferon alpha , interferon beta , and c-jun genes . Early-passage clone 7 cells exhibited chromosome 5 monosomy , while late-passage cells contained one normal chromosome 5 and a derivative ( 5q12q ) . Southern analysis of the three cell lines revealed no consistent loss of loci for the interferon and c-jun genes , although early-passage clone 7 cells had one half the gene copy number for the interferon beta and c-jun genes and both early- and late-passage clone 7 cells lacked DNA sequences hybridizing with the probe for interferon alpha . CONCLUSION This pattern of passage-dependent spontaneous transformation of rat ovarian surface epithelial cells in vitro supports the hypothesis that repetitious ovulation contributes to the etiology of human ovarian cancer . OUTPUT: evading growth suppressors;sustaining proliferative signaling INPUT: It is crucial for organ homeostasis that epithelia have effective mechanisms to restrict motility and cell proliferation in order to maintain tissue architecture . On the other hand , epithelial cells need to rapidly and transiently acquire a more mesenchymal phenotype , with high levels of cell motility and proliferation , in order to repair epithelia upon injury . Cross talk between cell-cell and cell-matrix signaling is crucial for regulating these transitions . The Pak1-betaPIX-GIT complex is an effector complex downstream of the small GTPase Rac1 . We previously showed that translocation of this complex from cell-matrix to cell-cell adhesion sites was required for the establishment of contact inhibition of proliferation . In this study , we provide evidence that this translocation depends on cadherin function . Cadherins do not recruit the complex by direct interaction . Rather , we found that inhibition of the normal function of cadherin or Pak1 leads to defects in focal adhesion turnover and to increased signaling by phosphatidylinositol 3-kinase . We propose that cadherins are involved in regulation of contact inhibition by controlling the function of the Pak1-betaPIX-GIT complex at focal contacts . OUTPUT:

evading growth suppressors

HoC_dynamic_5_shot98

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: Identification of the proteins that are associated with estrogen receptor ( ER ) status is a first step towards better understanding of the hormone-dependent nature of breast carcinogenesis . Although a number of gene expression analyses have been conducted , protein complement has not been systematically investigated to date . Because proteins are primary targets of therapeutic drugs , in this study , we have attempted to identify proteomic signatures that demarcate ER-positive and -negative breast cancers . Using highly enriched breast tumor cells , replicate analyses from 3 ERα+ and 3 ERα- human breast tumors resulted in the identification of 2,995 unique proteins with ≥2 peptides . Among these , a number of receptor tyrosine kinases and intracellular kinases that are abundantly expressed in ERα+ and ERα- breast cancer tissues were identified . Further , label-free quantitative proteome analysis revealed that 236 proteins were differentially expressed in ERα+ and ERα- breast tumors . Among these , 141 proteins were selectively up-regulated in ERα+ , and 95 proteins were selectively up-regulated in ERα- breast tumors . Comparison of differentially expressed proteins with a breast cancer database revealed 98 among these have been previously reported to be involved in breast cancer . By Gene Ontology molecular function , dehydrogenase , reductase , cytoskeletal proteins , extracellular matrix , hydrolase , and lyase categories were significantly enriched in ERα+ , whereas selected calcium-binding protein , membrane traffic protein , and cytoskeletal protein were enriched in ERα- breast tumors . Biological process and pathway analysis revealed that up-regulated proteins of ERα+ were overrepresented by proteins involved in amino acid metabolism , proteasome , and fatty acid metabolism , while up-regulated proteins of ERα- were overrepresented by proteins involved in glycolysis pathway . The presence and relative abundance of 4 selected differentially abundant proteins ( liprin-α1 , fascin , DAP5 , and β-arrestin-1 ) were quantified and validated by immunohistochemistry . In conclusion , unlike in vitro cell culture models , the in vivo signaling proteins and pathways that we have identified directly from human breast cancer tissues may serve as relevant therapeutic targets for the pharmacological intervention of breast cancer . OUTPUT: cellular energetics INPUT: Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers . A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation . We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products . This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine ( NMBzA ) , and the response that increased levels of dietary vitamin E has on this induced carcinogenesis . Female C57BL/6 mice received NMBzA or vehicle ( corn oil ) i.p. weekly for 3 weeks . Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet . As an assessment of free radical activity , exhaled ethane was measured prior to killing the animals at 26 weeks . Esophagi from the various mice groups were assessed for size and frequency of tumors . Livers homogenates were analyzed for vitamins A and E , lipid fluorescence , conjugated dienes and malondialdehyde . Hepatic levels of vitamin A and E were decreased ( P &lt ; 0.05 ) and indices of lipid peroxidation were greater ( P &lt ; 0.05 ) in NMBzA-treated mice relative to controls . Lipid peroxidation and serum transaminases ( ALT and AST ) were greatest in mice given NMBzA and infected with the retroviruses . Incidence of esophageal tumors were also greatest in the NMBzA-treated , immunocompromised animals . Mice fed vitamin E-supplemented diets showed increased ( P &lt ; 0.05 ) hepatic concentrations of vitamin E and vitamin A , decreased activities of serum transaminases , decreased indices of lipid peroxidation , and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice . These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals . OUTPUT: avoiding immune destruction INPUT: BACKGROUND Folate ( vitamin B9 ) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate ( dTMP ) and s-adenosylmethionine ( AdoMet ) . The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance , but still unclear . We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis , which is prominent in prostate cells . We , therefore , hypothesized that prostate cells might be highly susceptible to genetic , epigenetic and phenotypic changes consequent to folate restriction . RESULTS We studied the consequences of long-term , mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate ( TRAMP ) model , recapitulating different stages of prostate cancer ; benign , transformed and metastatic . High-performance liquid chromatography analysis demonstrated that mild folate depletion ( 100 nM ) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines . Random oligonucleotide-primed synthesis ( ROPS ) revealed a significant increase in uracil misincorporation and DNA single strand breaks , while spectral karyotype analysis ( SKY ) identified five novel chromosomal rearrangements in cells grown with mild folate depletion . Using global approaches , we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine , indicating a broad effect of folate depletion on epigenetic regulation . These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion . CONCLUSIONS This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate ( 2 microM ) routinely used in tissue culture . In addition , we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells . These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo , where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches . OUTPUT: genomic instability and mutation INPUT: Metastases in the sellar region are rare and are frequently found incidentally or in necropsies . Only 7% are reported to be symptomatic . Diabetes insipidus , anterior pituitary dysfunction , visual field defects , headache/pain and ophthalmoplegia are the most commonly reported symptoms . We present the cases of two male patients with a small-cell lung carcinoma whose first clinical symptoms were due to pituitary metastasis . One case presented with symptoms of cavernous sinus invasion and panhypopituitarism and the other case with diabetes insipidus . Both patients had a rapid progression of their disease despite chemotherapy and died after a few months . Pituitary metastases occur most commonly with breast cancer in women and lung cancer in men . The presence of polyuria and polydipsia in an oncologic patient should alert the physician for diabetes insipidus and , if confirmed , an imaging procedure of the pituitary gland is mandatory . Treatment for these tumors is often multimodal and includes surgery , radiation therapy , chemotherapy and hormone replacement . Although surgical series have not shown any significant survival benefits given by tumor resection , the patient's quality of life may be improved . OUTPUT: activating invasion and metastasis INPUT: Ovarian hormones have a pivotal role in the control of proliferation in the mammary gland , and cumulative life-time exposure to ovarian hormones is known to be a determinant of breast cancer risk . We have shown previously that a p.o.-active , long-acting butyrate analogue , sodium phenylbutyrate ( PB ) , reduced proliferation in normal and malignant human breast cells in culture and reduced expression of ovarian hormone receptors , suggesting that PB had cellular effects consistent with decreasing breast cancer risk . The aim of this study was to determine the in vivo effects of PB in the normal mammary gland on epithelial cell proliferation , estrogen receptor alpha ( ER alpha ) expression , and cyclin D1 expression . BALB/c mice were treated with PB , delivered by mini-osmotic pumps , for 7 days . Moderate ( 250 mg/kg/day ) and high ( 500 mg/kg/day ) PB treatment resulted in a decrease in proliferation in mammary epithelial cells ( P &lt ; 0.001 ) , determined by bromodeoxyuridine incorporation . Analysis of ER alpha immunostaining revealed a significant reduction in moderate- and high-treatment groups ( P = 0.01 and P = 0.02 ) , and expression of cyclin D1 was virtually ablated ( P &lt ; 0.001 ) . Histone deacetylase inhibition is a known mechanism of butyrate action , and consistent with this , PB increased levels of acetylated histone H3 in the mammary gland . In summary , PB decreased proliferation in the mammary gland in vivo at clinically achievable doses . Decreased proliferation was accompanied by changes in the levels of ER alpha and cyclin D1 . These data show that PB modulates parameters thought to be involved in the carcinogenic process in the normal mammary gland , and compounds in this class may therefore be useful candidates for breast cancer chemoprevention . OUTPUT: sustaining proliferative signaling INPUT: Previous studies with selenium and/or vitamin E in prostate carcinogenesis animal models have been negative , but these models may not involve oxidative stress mechanisms . In this study , we examined the potential of selenomethionine and alpha-tocopherol to modulate prostate cancer development in the testosterone plus estradiol-treated NBL rat , a model that does involve sex hormone-induced oxidative stress mechanisms and prostatic inflammation . One week following the implantation with hormone-filled Silastic implants , rats were fed diets containing l-selenomethionine ( 1.5 or 3.0 mg/kg ) , DL-alpha-tocopherol acetate ( 2,000 or 4,000 mg/kg ) , or a natural ingredient control diet ( NIH-07 ) . The development of prostate carcinomas was not affected by dietary treatment with either agent . Food intake , body weight , and mortality were also not affected . The high dose of selenomethionine reduced the severity of epithelial dysplasia in the lateral prostate that was not associated with inflammation , and alpha-tocopherol reduced in a dose-related fashion the incidence of marked inflammation and marked epithelial dysplasia in the lateral prostate , regardless of whether these lesions were associated with inflammation. alpha-Tocopherol significantly increased the incidence of adenocarcinomas of the mammary glands at both dietary concentrations . Collectively , our findings suggest that selenomethionine and alpha-tocopherol supplementation does not prevent prostate cancer in rats fed diets with nutritionally adequate levels of selenium and vitamin E. Importantly , the results of the current animal studies and those reported previously were fully predictive of the outcome of the Selenium and Vitamin E Cancer Prevention Trial . OUTPUT:

tumor promoting inflammation

HoC_dynamic_5_shot99

***TASK*** The task is to perform a semantic classification of the article according to the hallmarks of cancer based on its abstract. ***INPUT*** The input is an abstract text. ***DOCUMENTATION*** There are 10 cancer hallmarks you will need to decide whether the article is related to, including: activating invasion and metastasis sustaining proliferative signaling resisting cell death cellular energetics genomic instability and mutation evading growth suppressors inducing angiogenesis enabling replicative immortality avoiding immune destruction tumor promoting inflammation ***OUTPUT*** The output should be topics from the above 10 topics, that are related to the input article. Please note one article can be related to multiple topics. Output format: provide a semicolon-separated list of relevant topics. ***EXAMPLES*** INPUT: BACKGROUND Death receptors ( DR ) of the TNF family function as anti-tumor immune effector molecules . Tumor cells , however , often exhibit DR-signaling resistance . Previous studies indicate that radiation can modify gene expression within tumor cells and increase tumor cell sensitivity to immune attack . The aim of this study is to investigate the synergistic effect of sub-lethal doses of ionizing radiation in sensitizing colorectal carcinoma cells to death receptor-mediated apoptosis . METHODOLOGY/PRINCIPAL FINDINGS The ability of radiation to modulate the expression of multiple death receptors ( Fas/CD95 , TRAILR1/DR4 , TRAILR2/DR5 , TNF-R1 and LTβR ) was examined in colorectal tumor cells . The functional significance of sub-lethal doses of radiation in enhancing tumor cell susceptibility to DR-induced apoptosis was determined by in vitro functional sensitivity assays . The longevity of these changes and the underlying molecular mechanism of irradiation in sensitizing diverse colorectal carcinoma cells to death receptor-mediated apoptosis were also examined . We found that radiation increased surface expression of Fas , DR4 and DR5 but not LTβR or TNF-R1 in these cells . Increased expression of DRs was observed 2 days post-irradiation and remained elevated 7-days post irradiation . Sub-lethal tumor cell irradiation alone exhibited minimal cell death , but effectively sensitized three of three colorectal carcinoma cells to both TRAIL and Fas-induced apoptosis , but not LTβR-induced death . Furthermore , radiation-enhanced Fas and TRAIL-induced cell death lasted as long as 5-days post-irradiation . Specific analysis of intracellular sensitizers to apoptosis indicated that while radiation did reduce Bcl-X(L) and c-FLIP protein expression , this reduction did not correlate with the radiation-enhanced sensitivity to Fas and/or TRAIL mediated apoptosis among the three cell types . CONCLUSIONS/SIGNIFICANCE Irradiation of tumor cells can overcome Fas and TRAIL resistance that is long lasting . Overall , results of these investigations suggest that non-lethal doses of radiation can be used to make human tumors more amenable to attack by anti-tumor effector molecules and cells . OUTPUT: resisting cell death INPUT: The cellular mechanisms of anti-androgen-induced tumor regression have not been investigated in great detail . We have compared the induction of cell death in the androgen-dependent , non-invasive LNCaP prostate cancer cell line by Casodex and TNF-alpha . Both agents induce a dose and time-dependent decrease in cell viability in vitro . However , Casodex does not induce classical DNA fragmentation to oligonucleosomes typically induced by TNF-alpha , but rather induces cleavage to form intermediate 60 kb DNA fragments . RT-PCR based analysis demonstrates that in LNCaP cells Casodex coordinately alters the expression of steady-state level of mRNAs of several matrix metalloproteases and their cognate inhibitors ( most notably MMP2 and TIMP1 ) . Zymography and reverse zymography confirm that the ratio of metalloprotease(s) to inhibitor(s) is altered in favor of activation of the proteases . In a small percentage of the treated LNCaP cells , the activation of the extracellular matrix ( ECM)-proteases by Casodex also induces an invasive phenotype . The acquisition of an invasive phenotype is not seen when LNCaP cells are treated with TNF-alpha , and is not seen when the LNCaP cells are treated with both compounds simultaneously , suggesting that the phenomenon may be specific to particular classes of compounds . These observations have significant implications in the treatment of prostate cancer , since the appearance of a more aggressive phenotype following treatment is clearly undesirable . OUTPUT: resisting cell death;activating invasion and metastasis INPUT: INTRODUCTION Necrosis at the tumor center is a common feature of aggressive breast cancers and has been associated with poor prognosis . It is commonly identified by means of invasive histopathology , which often correlates with morbidity and potential tumor cell dissemination , and limits the reconstruction of the whole necrotic domain . In this study we hypothesized that non covalent association to serum albumin ( SA ) and covalent binding to ligands for tumor-abundant cell receptors should synergistically drive selective accumulation and prolonged retention of imaging and therapeutic agents in breast tumor necrotic domains enabling in vivo identification , imaging and possibly treatment of such tumors . METHODS Cyclo-Arg-Gly-Asp-D-Phe-Lys ( c(RGDfK) ) were conjugated to bacteriochlorophyll-derivatives ( Bchl-Ds ) , previously developed as photodynamic agents , fluorescent probes and metal chelators in our lab . The c(RGDfK) component drives ligation to alphaVbeta3 integrin receptors over-expressed by tumor cells and neo-vessels , and the Bchl-D component associates to SA in a non-covalent manner . STL-6014 , a c(RGDfK)-Bchl-D representative , was i.v. injected to CD-1 , nude female mice bearing necrotic and non-necrotic human MDA-MB-231-RFP breast cancer tumors . The fluorescence signals of the Bchl-Ds and RFP were monitored over days after treatment , by quantitative whole body imaging and excised tumor/tissue samples derived thereof . Complementary experiments included competitive inhibition of STL-6014 uptake by free c(RGDfK) , comparative pharmacokinetics of nonconjugated c(RGDfK) Bchl-D ( STL-7012 ) and of two human serum albumin ( HSA ) conjugates : HSA-STL-7012 and HSA-STL-6014 . RESULTS STL-6014 and STL-7012 formed complexes with HSA ( HSA/STL-6014 , HSA/STL-7012 ) . STL-6014 , HSA-STL-7012 and HSA-STL-6014 , selectively accumulated at similar rates , in tumor viable regions over the first 8 h post administration . They then migrated into the necrotic tumor domain and presented tumor half lifetimes ( T1/2 ) in the range of days where T1/2 for HSA-STL-6014 &gt ; STL-6014 &gt ; HSA-STL-7012 . No accumulation of STL-7012 was observed . Pre-injection of c(RGDfK) excess , prevented the uptake of STL-6014 in the small , but not in the large tumors . CONCLUSIONS Non-covalent association to SA and covalent binding to c(RGDfK) , synergistically enable the accumulation and prolonged retention of Bchl-Ds in the necrotic regions of tumors . These findings provide novel guidelines and strategy for imaging and treatment of necrotic tumors . OUTPUT: resisting cell death INPUT: Chronic inflammation is a risk factor for the development of colon cancer , providing genotoxic insults , growth and pro-angiogenic factors that can promote tumorigenesis and tumor growth . Immunomodulatory agents can interfere with the inflammation that feeds cancer , but their impact on the transformed cell is poorly understood . The calcium/calcineurin signaling pathway , through activation of NFAT , is essential for effective immune responses , and its inhibitors cyclosporin A ( CsA ) and FK506 are used in the clinics to suppress immunity . Moreover , the kinases GSK3β and mTOR , modulated by PI-3K/Akt , can inhibit NFAT activity , suggesting a cross-talk between the calcium and growth factor signaling pathways . Both NFAT and mTOR activity have been associated with tumorigenesis . We therefore investigated the impact of calcineurin and PI-3K/mTOR inhibition in growth of human colon carcinoma cells . We show that despite the efficient inhibition of NFAT1 activity , FK506 promotes tumor growth , whereas CsA inhibits it due to a delay in cell cycle progression and induction of necroptosis . We found NFκB activation and mTORC1 activity not to be altered by CsA or FK506 . Similarly , changes to mitochondrial homeostasis were equivalent upon treatment with these drugs . We further show that , in our model , NFAT1 activation is not modulated by PI3K/mTOR . We conclude that CsA slows cell cycle progression and induces necroptosis of human carcinoma cell lines in a TGFβ- , NFAT- , NFκB- and PI3K/mTOR-independent fashion . Nevertheless , our data suggest that CsA , in addition to its anti-inflammatory capacity , may target transformed colon and esophagus carcinoma cells without affecting non-transformed cells , promoting beneficial tumoristatic effects . OUTPUT: sustaining proliferative signaling;tumor promoting inflammation INPUT: Liver cancer , predominantly hepatocellular carcinoma ( HCC ) , represents a complex and fatal malignancy driven primarily by oxidative stress and inflammation . Due to dismal prognosis and limited therapeutic intervention , chemoprevention has emerged as a viable approach to reduce the morbidity and mortality of HCC . Pomegranate fruit is a rich source of phytochemicals endowed with potent antioxidant and anti-inflammatory properties . We previously reported that pomegranate phytochemicals inhibit diethylnitrosamine ( DENA)-initiated hepatocarcinogenesis in rats though nuclear factor E2-related factor 2 ( Nrf2)-mediated antioxidant mechanisms . Since Nrf2 also acts as a key mediator of the nuclear factor-kappaB ( NF-κB)-regulated inflammatory pathway , our present study investigated the anti-inflammatory mechanisms of a pomegranate emulsion ( PE ) during DENA-induced rat hepatocarcinogenesis . Rats were administered with PE ( 1 or 10 g/kg ) 4 weeks before and 18 weeks following DENA initiation . There was a significant increase in hepatic expressions of inducible nitric oxide synthase , 3-nitrotyrosine , heat shock protein 70 and 90 , cyclooxygenase-2 and NF-κB in DENA-exposed rat livers . PE dose-dependently suppressed all aforementioned elevated inflammatory markers . A conspicuous finding of this study involves lack of cardiotoxicity of PE as assessed by monitoring cardiac function using noninvasive echocardiography . Our results provide substantial evidence that suppression of the inflammatory cascade through modulation of NF-κB signaling pathway may represent a novel mechanism of liver tumor inhibitory effects of PE against experimental hepatocarcinogenesis . Data presented here coupled with those of our earlier study underline the importance of simultaneously targeting two interconnected molecular circuits , namely , Nrf2-mediated redox signaling and NF-κB-regulated inflammatory pathway , by pomegranate phytoconstituents to achieve chemoprevention of HCC . OUTPUT: tumor promoting inflammation INPUT: BACKGROUND Non-alcoholic fatty liver disease ( NAFLD ) is associated with obesity , insulin resistance and hepatic steatosis . Non-alcoholic steatohepatitis ( NASH ) is a serious consequence of NAFLD where chronic tissue damage and inflammation result in fibrosis which may progress to cirrhosis . Transforming growth factor beta1 ( TGFbeta1 ) , proinflammatory cytokines and oxidative stress are thought to play crucial roles in the pathogenesis of these conditions . The contributions of individual liver cell types to fibrogenesis remain controversial and the influence of selenium status has not been investigated . METHODS In this study we have used a cell culture model of fat-loading using oleate-treated human hepatoblastoma ( C3A ) cells to investigate how fat-loading and selenium status might influence the production of collagen in response to TGFbeta1 . The secretion of inflammatory cytokines was also investigated , together with the epithelial character of the treated cells . RESULTS We found that in response to treatment with TGFbeta1 , C3A cells produced mRNA encoding the pro-alphaI chain of procollagen I , secreted procollagen I peptide , up-regulated production of the proinflammatory cytokine interleukin-8 ( IL-8 ) and the mesenchymal marker vimentin , and down-regulated albumin production . Most of these responses were considerably enhanced when cells were fat-loaded with oleate and were attenuated by selenium addition at a dose that optimised the expression of thioredoxin reductase and glutathione peroxidase . CONCLUSIONS Our data establish that both fat-loading and suboptimal selenium status enhance collagen and IL-8 production by C3A hepatocytes in response to TGFbeta1 , possibly as part of an epithelial to mesenchymal transition . GENERAL SIGNIFICANCE These findings suggest that the hepatocyte may be an important contributor to the pathogenesis of fibrosis associated with NAFLD . OUTPUT:

tumor promoting inflammation